KR100581163B1 - A hybrid Bacillus thuringiensis toxin, a nucleic acid encoding the same and a method for controlling an insect with the same - Google Patents

Abstract

Synthetic nucleotide sequences optimized for expression in plants encode various forms of hybrid Bacillus thuringiensis delta-endotoxin H04 in which the toxin moiety contains domains I and II of Cry1Ab and domain III of Cry1C. Compositions and formulations containing such pesticidal toxins can control insect pests. Further, the present invention relates to a method for producing a hybrid toxin and a method for controlling insect pests by using the nucleotide sequence in, for example, microorganisms and endowing insect resistance with transgenic plants.

Hybrid Bacillus thuringiensis toxin, transformation, chimeric gene, insecticidal toxin

Description

Hybrid Bacillus thuringiensis toxin, a nucleic acid encoding the same and a method for controlling an insect with the same}

The present invention provides insecticidal toxins derived from Bacillus thuringiensis insecticidal determinant proteins, nucleic acid sequences that express and produce the toxins, and methods for preparing and using the toxins and corresponding nucleic acid sequences to control pests. It is about.

Pests are a major source of crop loss. In the United States alone, trillions of dollars are lost every year due to infections with various insects. In addition to the loss of field crops, pests also burden gardeners and homeowners by burdening vegetable and fruit growers and ornamental flower producers.

Pests are controlled primarily by the intensive application of active chemical insecticides through inhibition of insect growth, prevention of insect nutrition or reproduction, or death of insects. Thus, good insect control can be achieved, but the chemical can sometimes affect other worms. Another problem resulting from the widespread use of chemical pesticides is the emergence of resistant insect strains. This has been partially alleviated by various resistance control strategies, but there is an increasing demand for alternative pest control agents.

Biological insect control agents such as Bacillus thuringiens strains expressing insecticidal toxins have been applied to provide alternatives or supplements to chemical insecticides with satisfactory results. Bacillus thuringiensis belongs to a large group of Gram-positive aerobic spore-forming bacteria. ratio. B. cereus or b. Unlike other closely related Bacillus species, such as B. anthracis, most of the Bacillus thuringiens species known so far are due to their crystal structure during the sporulation process and are generally referred to as crystals. To produce. The crystal consists of a pesticidally active crystalline protoxin protein, the so-called δ-endotoxin. These protein crystals cause toxicity to insects of Bacillus thuringiensis. δ-endotoxins do not exhibit pesticidal activity until after crystals have been orally ingested until the crystals dissolve in the gastric juice of the target insect. In most cases, the actual toxic components are released from the pretoxins as a result of proteolytic cleavage caused by the protease action from the insect's digestive tract. The δ-endotoxins of the various Bacillus thuringiens strains are characterized by high specificity with respect to certain target insects, in particular various poultry, superpod and fly larvae and high activity against these larvae. A further advantage when using the δ-endotoxin of Bacillus thuringiens is that the toxin is harmless to humans, other mammals, birds and fish.

Based on sequence homology and pesticidal specificity, Bacillus thuringiensis crystalline proteins have been classified into different classes. The Cry1 family of proteins, produced as 140kDa protoxins and active against the eyes, are best studied. The mode of action of crystalline proteins has been described to some extent. After oral ingestion, the crystals dissolve in the alkaline environment of the larva's mesenteric. The solubilized protein is then treated with a mesenchymal proteinase (such as trypsin) to about 65 kDa proteinase-resistant toxic fragment that binds to receptors on epithelial cells of the insect mesent and penetrates the cell membrane. This eventually leads to cell rupture and death of the larvae.

The activity spectrum of certain crystalline proteins is measured to a significant extent by receptor development on mesenteric epithelial cells of sensitive insects. Spectra are co-measured by the efficiency of solubilization of the crystalline protein and its proteolytic activation in vivo. The importance of the binding of the crystalline protein to the mesenteric epithelial receptor is further evidenced that in insects with advanced resistance to one of the crystalline proteins, the binding of the crystalline protein to the mesenteric epithelial cells of the resistant insect is significantly reduced.

Over the past years, genes encoding some of these deterministic proteins have been isolated and their expression in heterologous hosts has been shown to provide another tool for the control of economically important pests. In particular, the expression of pesticidal toxins in transgenic plants, such as Bacillus thuringiensis crystalline protein, provided effective protection against selected pests and commercialized transgenic plants expressing these toxins reduced farmers' application of chemical insect control agents. I could make it. It is also possible to express recombinant toxins with the selected combination of functions designed to increase the degree of pesticidal activity against a particular insect or insect species or to broaden the spectrum of insects in which the toxin protein is active. For example, a chimeric insecticidal protein having a novel sequence not found in nature can be constructed by combining a toxin portion from one δ-endotoxin and a different pretoxin (tail) portion of a different δ-endotoxin. Document: WO 98/15170, which is incorporated herein by reference.

Toxic fragments of crystalline proteins are believed to consist of three separate structural domains. Domain I, the most N-terminal domain, consists of seven α-helices and may be partially or wholly inserted into the target cell membrane. Domain II comprises 3 β-sheets in the so-called lattice-stereoform. Most investigators believe that domain II interacts with the receptor, thereby determining toxin specificity. In addition, there is a great deal of evidence associated with domain II residues in specific toxicity and high affinity binding. Domain III, the most C-terminal domain, consists of two β-sheets in the so-called jellyroll conformation and also determines specificity. For example, specific activity can be changed by exchanging domain III between toxins by in vivo recombination between coding regions. Binding experiments using these hybrids suggest that Domain III is involved in binding to putative receptors of target insects, suggesting that Domain III may play a role in specificity through its role in receptor recognition. In light of the Cry1 sequence, domain I corresponds to amino acid residues 28 to 260, domain II to about 260 to 460, and domain III to about 460 to 600 (see Nakamura et al. , Agric. Biol. Chem. 54 (3): 715-724 (1990); Li et al., Nature 353: 815-821 (1991); Ge et al., J. Biol. Chem. 266 (27): 17954-17958 (1991); and Honee et al., Mol. Microbiol. 5 (11): 2799-2806 (1991); Each is incorporated herein by reference]. US Pat. No. 5,736,131, which is incorporated herein by reference, contains a Bacillus thuringiens hybrid toxin fragment comprising domain III of the first Cry protein at the C-terminus and domains I and II of the second Cry protein at the N-terminus. This is described. Such hybrid crystal proteins have altered pesticidal specificity. See, eg, De Maagd et al., Appl. Environ Microbiol. 62 (5): 1537-1543 (1996) also includes domains I and II of Cry1Ab at the N-terminus and domain III of Cry1C at the C-terminus. Reportedly, H04 is highly toxic to Spodoptera exigua compared to the parent Cry1Ab toxin and more toxic to the Cry1C parent toxin [Bosch et al., FEMS Microbiology Letters 118: 129]. -134 (1994); Bosch et al., Bio / Technology 12: 915-918 (1994); De Maagd et al., Appl. Environ. Microbiol. 62 (8): 2753-2757 (1996); And De Maagd et al., Mol. Microbiol. 31 (2): 463-471 (1999); Each is incorporated herein by reference].

Despite the previous success achieved with the incorporation of insect resistance genes through breeding programs and genetic engineering, the need for finding new and effective insect control agents is urgent but not realized. In particular, there is a need for control agents that target economically important pests such as European Corn Borer and Fall Army Worm and effectively control insect species that are resistant to current insect control agents. Also preferred are formulations that are applied to minimize the burden on the environment.

The present invention fulfills these needs by providing novel genes encoding hybrid Bacillus thuringiensis toxins, including synthetic nucleotide sequences optimized for expression in plants. In a preferred embodiment, the new gene sequence encodes various forms of the hybrid Bacillus thuringiensis delta-endotoxin H04 wherein the toxin moiety contains domains I and II of Cry1Ab and domain III of Cry1C. The hybrid Bacillus thuringiensis toxin encoded by this new gene is responsible for economically important pests such as chestnut moth, pink bollworm, tobacco budworm, European moth and diamondback moth. Highly active. Hybrid Bacillus thuringiensis toxins can be used in many insect control strategies to achieve maximum efficiency with minimal impact on the environment.

The present invention further provides hybrid insecticidal toxins produced by the expression of the nucleotide sequences of the present invention, and those hybrid insecticidals that can inhibit the ability of pests to survive, grow or reproduce or limit damage or loss associated with insects in crops. It relates to compositions and formulations containing toxins. In addition, the present invention provides a method for preparing a hybrid toxin and a method for conferring insecticidal resistance using the nucleotide sequence in a transformed plant, for example, and a method for using the toxin and a composition and formulation containing the toxin, for example. For example, the present invention relates to methods of applying such toxins, compositions or formulations to insect infected areas, insect sensitive areas or plants to be treated prophylactically to confer protection or resistance to harmful insects. Hybrid toxins can be used in many insect control strategies to achieve maximum efficiency with minimal impact on the environment.

According to one aspect, the present invention comprises delivering to an insect an effective amount of a hybrid Bacillus thuringiensis toxin comprising domains I and II from the Cry1Ab toxin bound in the amino to carboxy direction to domain III from the Cry1C toxin And insects selected from the group consisting of chestnut moth, pink cotton worm, tobacco moth, European light moth and Chinese cabbage moth. In a preferred embodiment, the hybrid Bacillus thuringiensis toxin comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 2, 4, 6, 8 or 10. In a more preferred embodiment, the hybrid Bacillus thuringiensis toxin comprises SEQ ID NO: 2, 4, 6, 8 or 10.

In another embodiment of the above described method of the invention, the hybrid Bacillus thuringiensis toxin comprises a C-terminal tail region, such as a Cry1C tail region or a Cry1Ab tail region. The C-terminal tail region can be full length or can be cleaved, for example, to about 40 amino acids in length.

In a preferred embodiment of the above-described method of the invention, delivering an effective amount of a hybrid Bacillus thuringiensis toxin to an insect is transformed into a recombinant DNA sequence comprising a nucleotide sequence encoding the hybrid Bacillus thuringiensis toxin to the insect. Feeding transgenic plant tissues or contacting insects thereto, wherein expression of the hybrid Bacillus thuringiensis toxin in the transgenic plant tissue confers resistance to insects. Preferably, the nucleotide sequence is substantially identical to SEQ ID NO: 1, 3, 5, 7 or 9.

According to another aspect, the invention provides an N-terminal toxin moiety comprising (a) domains I and II from Cry1Ab toxin bound in amino to carboxy direction to domain III from Cry1C toxin and (b) from Cry1Ab toxin An isolated nucleic acid molecule is provided comprising a nucleotide sequence encoding a hybrid Bacillus thuringiensis toxin comprising a C-terminal tail region. Preferably, the hybrid Bacillus thuringiensis toxin comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 6, 8, or 10. More preferably, the hybrid Bacillus thuringiensis toxin comprises SEQ ID NO: 6, 8 or 10. Even more preferably, the nucleotide sequence is at least 90% identical to SEQ ID NO: 5, 7 or 9. Most preferably, the nucleotide sequence comprises SEQ ID NO: 5, 7 or 9.

The present invention provides a chimeric gene comprising a heterologous promoter sequence operably linked to a nucleic acid molecule of the invention as described above; A recombinant vector comprising the chimeric gene; Transgenic host cells (eg, plant cells) comprising said chimeric genes; Transgenic plants (eg, corn, cotton, rice or cabbage plants) comprising said transgenic plant cells; And seed from the transgenic plant.

According to another aspect, the present invention provides a nucleic acid promoter sequence that promotes (a) elevated levels of transcription of an associated coding sequence in a plant, and (b) a nucleic acid molecule according to the invention that is operably linked to said promoter sequence. A method of protecting a plant from insects, including the expression of a hybrid Bacillus thuringiensis toxin in a plant transformed with a chimeric gene comprising, wherein the expression of the hybrid Bacillus thuringiensis toxin in the plant protects the plant from insects. Provide a way to.             

According to another aspect, the present invention provides a hybrid Bacillus toxin that is active against an insect by (a) obtaining a transformed host cell according to the present invention and (b) expressing the nucleic acid molecule of the present invention in said transformed host cell. It provides a method for producing a hybrid Bacillus thuringiensis toxin active against insects, including the step of obtaining the same.

According to another aspect, the present invention provides a method for producing a plant resistant to insects, comprising introducing a nucleic acid molecule according to the present invention into a plant, wherein the nucleic acid molecule controls insects in the plant. Provided are methods that can be expressed in an effective amount.

According to another aspect, the invention provides an isolated nucleic acid molecule comprising SEQ ID NO: 3, 5, 7, 9, 11, 12, 13, 14, 15, 16 or 17; Chimeric genes comprising heterologous promoter sequences operably linked to such nucleic acid molecules; Recombinant vectors comprising such chimeric genes; Transgenic host cells (eg, plant cells) comprising such chimeric genes; Transgenic plants (eg, corn, cotton, rice or cabbage plants) comprising such transgenic plant cells; And seeds derived from such transgenic plants.

According to a further aspect, the present invention relates to an N-terminal toxin moiety comprising (a) domains I and II from Cry1Ab toxin bound amino to carboxy to domain III from Cry1C toxin and (b) from Cry1Ab toxin A hybrid Bacillus thuringiensis toxin comprising a C-terminal tail region is provided. Preferably, the hybrid Bacillus thuringiensis toxin of the present invention comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 6, 8 or 10. More preferably, the hybrid Bacillus thuringiensis toxin of the present invention comprises SEQ ID NO: 6, 8 or 10.

According to a further aspect, the present invention provides a composition comprising the hybrid Bacillus thuringiensis toxin of the present invention in an amount effective to control insects.

Other aspects and advantages of the invention will be apparent to those skilled in the art from the following description of the invention and the study of non-limiting examples.

Brief description of the sequences in the sequence listing

SEQ ID NO: 1 has a toxin moiety comprising domains I and II of Cry1Ab at the N-terminus and domain III of Cry1C at the C-terminus. De Maagd et al., Appl. Environ. Microbiol. 62 (5): 1537-1543 (1996), presents a nucleotide sequence encoding the H04 hybrid toxin and full-length Cry1C tail portion.

SEQ ID NO: 2 shows the amino acid sequence of the H04 hybrid toxin encoded by the nucleotide sequence described in SEQ ID NO: 1, comprising the full length Cry1C tail portion together with toxin domains I and II of Cry1Ab and toxin domain III of Cry1C.

SEQ ID NO: 3 presents a synthetic nucleotide sequence that encodes a toxin portion of H04 that does not contain a tail as the trypsin site is cleaved.

SEQ ID NO: 4 sets forth the amino acid sequence of the H04 toxin moiety encoded by the synthetic nucleotide sequence set forth in SEQ ID NO: 3.

SEQ ID NO: 5 shows a synthetic nucleotide sequence encoding the toxin portion of H04 and the full length Cry1Ab tail portion.

SEQ ID NO: 6 shows the amino acid sequence of the H04 + Cry1Ab tail encoded by the synthetic nucleotide sequence set forth in SEQ ID NO: 5.

SEQ ID NO: 7 shows another synthetic nucleotide sequence that encodes a toxin portion of H04 and a full length Cry1Ab tail portion.

SEQ ID NO: 8 shows the amino acid sequence of the H04 + Cry1Ab tail encoded by the synthetic nucleotide sequence set forth in SEQ ID NO: 7.

SEQ ID NO: 9 shows a synthetic nucleotide sequence that encodes the first 40 amino acids of the toxin portion and Cry1Ab tail portion of H04.

SEQ ID NO: 10 shows the amino acid sequence of a Cry1Ab tail truncated to H04 + 40 amino acids encoded by the synthetic nucleotide sequence set forth in SEQ ID NO: 9.

SEQ ID NO: 11 shows the nucleotide sequence of construct pNOV1308 containing a constitutive corn ubiquitin promoter operably linked to a synthetic nucleotide sequence that encodes a toxin portion of H04 that does not include a tail as set forth in SEQ ID NO: 3.

SEQ ID NO: 12 shows the nucleotide sequence of construct pNOV1436, which contains a preferred corn MTL promoter in the roots, operably linked to a synthetic nucleotide sequence encoding the toxin portion of H04 and the full length Cry1Ab tail portion as set forth in SEQ ID NO: 5.

SEQ ID NO: 13 shows the nucleotide sequence of construct pNOV1441 containing a constitutive corn ubiquitin promoter operably linked to a synthetic nucleotide sequence encoding the toxin portion of H04 and the full length Cry1Ab tail portion as set forth in SEQ ID NO: 5.

SEQ ID NO: 14 shows the nucleotide sequence of construct pNOV1305, which contains a constitutive corn ubiquitin promoter operably linked to a synthetic nucleotide sequence encoding a toxin portion of H04 and a full length Cry1Ab tail portion as set forth in SEQ ID NO: 7.

SEQ ID NO: 15 shows the nucleotide sequence of construct pNOV1313, which contains a constitutive corn ubiquitin promoter operably linked to a synthetic nucleotide sequence encoding a toxin portion of H04 and a full length Cry1Ab tail portion as set forth in SEQ ID NO: 7.

SEQ ID NO: 16 shows the nucleotide sequence of construct pNOV1435, which contains a desired maize MTL promoter for roots, operably linked to the toxin portion of H04 as shown in SEQ ID NO: 9 and the synthetic nucleotide sequence encoding the first 40 amino acids of the Cry1Ab tail do.

SEQ ID NO: 17 shows a fragment of construct pZU578 containing an arabidopsis actin-2 promoter operably linked to the toxin portion of H04 as shown in SEQ ID NO: 9 and a synthetic nucleotide sequence encoding the first 40 amino acids of the Cry1Ab tail. The nucleotide sequence is shown.

Justice

"Activity" of the toxin of the present invention means that the toxin function as an orally active insect control agent may have a toxic effect or interrupt or interrupt insect nutrition. When the toxin of the present invention is delivered to an insect, the result is usually the killing of the insect or the insect does not consume a source that makes the toxin available to the insect.

"Associated / operably linked" means two nucleic acid sequences that are physically or functionally related. For example, a promoter or regulatory DNA sequence is a DNA sequence that encodes an RNA or protein when the two sequences are operatively linked or when the regulatory DNA sequence is placed in a situation that can affect the expression level of the coding or structural DNA sequence. Is referred to as "associated".

"Binding site" means a site on a molecule that is reversible such that the binding between site and toxicity is at least about 10 ⁴ dm ³ mole ^-1 .

A “chimeric gene” is a recombinant nucleic acid sequence in which a promoter or regulatory nucleic acid sequence encodes or associates with a nucleic acid sequence that encodes an mRNA or is expressed as a protein, such that the regulatory nucleic acid sequence may regulate transcription or expression of the associated nucleic acid sequence. to be. The regulatory nucleic acid sequences of the chimeric genes are not operably linked to normally associated nucleic acid sequences, as found in nature.

A "coding sequence" is a nucleic acid sequence that is transcribed into RNA such as mRNA, rRNA, tRNA, snRNA, sense RNA, or antisense RNA. Preferably, RNA is subsequently translated into an organism to produce a protein.             

Complementary: "Complementary" means two nucleotide sequences that include antiparallel nucleotide sequences that can pair with each other after hydrogen bonds are formed between complementary base residues in the antiparallel nucleotide sequence.

A "conservatively modified variant" of a particular nucleic acid sequence means a nucleic acid sequence that encodes the same or essentially the same amino acid sequence, or if the nucleic acid sequence does not encode an amino acid sequence, it means essentially the same sequence. Due to the degeneracy of the genetic code, many functionally identical nucleic acids encode any given polypeptide. For example, codons CGT, CGC, CGA, CGG, AGA and AGG all encode the amino acid arginine. Thus, at all positions where arginine is defined by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded protein. Such nucleic acid variations are "silent variations" which are a type of "conservatively modified variation". All nucleic acid sequences described herein that encode a protein also refer to all possible latent variations, unless stated otherwise. Those skilled in the art can appreciate that standard techniques can be used to modify each codon (except ATG, which is typically the only codon for methionine) to yield functionally identical molecules. Thus, each "potential variation" of a nucleic acid encoding a protein is inherent in each described sequence.

In addition, those skilled in the art will recognize that amino acids are chemically similar amino acids due to alterations in individual substitutions, deletions or additions that alter, add, or delete a small proportion (typically less than 5%, more typically less than 1%) of amino acids in the amino acid sequence It will be recognized that it is a "conservatively modified variant" substituted with. Conservative substitution tables that provide functionally similar amino acids are well known in the art. The following five groups each contain amino acids that are conservative substitutions for one another: aliphatic: glycine (G), alanine (A), valine (V), leucine (L), isoleucine (I); Aromatic: phenylalanine (F), tyrosine (Y), tryptophan (W); Sulfur-containing: methionine (M), cysteine (C); Basic: arginine (R), lysine (K), histidine (H); Acidic: Aspartic Acid (D), Glutamic Acid (E), Asparagine (N), Glutamine (Q) [Creighton (1984) Proteins, W.H. Freeman and Company]. In addition, individual substitutions, deletions or additions that alter, add or delete a single amino acid or a small proportion of amino acids in an encoded sequence are also "conservatively modified variations".

"Controlling" an insect means inhibiting the insect's ability to survive, grow, nourish and / or propagate or limit the damage or loss associated with the insect in the crop through a toxic effect. Although "controlling" an insect preferably means killing the insect, it may or may not mean killing the insect.

Corresponding: Within the scope of the present invention, "corresponds to" or "corresponds to" means that the nucleic acid coding sequences or amino acid sequences of the different δ-endotoxins of Bacillus thuringiensis are aligned at certain calculated positions when aligned with each other. Corresponding "nucleic acid or amino acid is a nucleic acid or amino acid that is aligned with said position but is not necessarily present at this exact numerical position relative to each nucleic acid coding sequence or amino acid sequence of a particular δ-endotoxin. Likewise, if the coding or amino acid sequence of a particular δ-endotoxin (eg Cry1B) is aligned with the coding or amino acid sequence of a reference δ-endotoxin (eg Cry1Ab), and with any calculated position of the Cry1Ab sequence, A nucleic acid or amino acid within a “corresponding” Cry1B sequence is a nucleic acid or amino acid that is aligned with the position of Cry1Ab but is not necessarily present at this exact numerical position of each nucleic acid coding sequence or amino acid sequence of the Cry1B toxin.

"Delivering" a toxin means contacting the toxin with an insect, resulting in toxic effects and control of the insect. Toxins can be expressed in a number of recognized ways, for example, by transgenic plant expression, formulated protein composition (s), sprayable protein composition (s), orally or by contact with insects, It may be delivered via a bait matrix or any other toxin delivery system known in the art.

As used herein, an "expression cassette" includes a nucleic acid sequence capable of inducing expression of said nucleotide sequence in a suitable host cell, including a promoter operably linked to a nucleotide sequence of interest operably linked to a termination signal. Means. Expression cassettes also typically include sequences required for proper translation of nucleotide sequences. An expression cassette comprising a nucleotide sequence of interest may be chimeric, meaning that one or more of the components of the expression cassette are heterologous to one or more of the other components of the expression cassette. In addition, expression cassettes may be naturally occurring but obtained in recombinant form useful for heterologous expression. Typically, however, expression cassettes are heterologous to the host. That is, the specific DNA sequence of the expression cassette is not naturally present in the host cell and must be introduced into the host cell or the progenitor of the host cell by the transformation process. Expression of the nucleotide sequence in the expression cassette may be under the control of a constitutive or inducible promoter that initiates transcription only when the host cell is exposed to some specific external stimulus. In the case of multicellular organisms such as plants, the promoter may also be specific for a particular tissue or organ or stage of development.

Gene: The term "gene" refers to any segment of DNA associated with a biological function and is used broadly. Thus, genes include coding sequences and / or regulatory sequences necessary for expression. In addition, genes include, for example, non-expressed DNA segments that form recognition sequences for other proteins. Genes can be obtained from a variety of sources, including cloning from a source of interest or synthesis from known or estimated sequence information, and can include sequences designed to have desirable parameters.

"Genetic genes of interest", when delivered to plants, are characterized by antibiotic resistance, viral resistance, insect resistance, disease resistance or resistance to other pests, herbicide content, improved nutritional value, improved efficiency or altered fertility in industrial processes. Any gene that imparts the same desirable properties. In addition, a "gene of interest" may be a gene that is delivered to a plant for production of commercially valuable enzymes or metabolites in the plant.

As used herein, "H04" refers to De Maagd et al., Wherein the toxin moiety comprises domains I and II of Cry1Ab at the N-terminus and domain III of Cry1C at the C-terminus. Appl. Environ. Microbiol. 62 (5): 1537-1543 (1996).

Heterologous Nucleic Acid Sequence: As used herein, the terms “heterologous nucleic acid [or DNA] sequence”, “exogenous nucleic acid [or DNA] segment” or “heterologous gene” each originate from a source that is foreign to a particular host cell or from the same source). When derived, it means a sequence modified from its original form. Thus, heterologous genes in host cells include genes that are endogenous to a particular host cell but that have been modified, for example, through the use of codon optimization. The term also encompasses a number of non-naturally occurring copies of naturally occurring sequences. Thus, the term refers to a nucleic acid segment that is exotic or heterologous to a cell, or is located at a location within a host cell nucleic acid that is homologous to the cell but whose elements are typically not found. Exogenous nucleic acid segments are expressed to yield exogenous peptides.

A “homologous” nucleic acid [or DNA] sequence is a nucleic acid [or DNA] sequence naturally associated with the host cell into which it is introduced.

"Homologous recombination" is the interchange of nucleic acid fragments between homologous nucleic acid molecules.

"Homoplastidic" means a plant, plant tissue or plant cell in which all of the pigment bodies are genetically identical; This is the normal state of the plant when the pigment is not transformed, mutated or genetically modified. In different tissues or developmental stages, the pigment is for example chloroplasts, chromosomes, etioplasts, amyloids and It may take other forms, such as colored bodies.

The term “identical” or% “identity” in the category of two or more nucleic acid or protein sequences, when compared and aligned for maximum correspondence, as determined by visual inspection or using a sequence comparison algorithm described below. , Or two or more sequences or substrings having the specified proportion of identical or identical amino acid residues or nucleotides.

"Pesticide" is defined as the toxic biological activity that can control insects, preferably kill them.

A nucleic acid sequence is "isocoding" with the nucleic acid sequence when the nucleic acid sequence encodes a polypeptide having the same amino acid sequence as the polypeptide encoded by the reference nucleic acid sequence.

A "isolated" nucleic acid molecule or an isolated enzyme is a nucleic acid molecule or enzyme that is not a natural product by being present away from its natural environment by a human hand. Isolated nucleic acid molecules or enzymes may be present in purified form or may be present in non-natural environments, such as, for example, recombinant host cells.

The "junction" between the toxin domains in the hybrid toxin, ie between the domains II and III of the hybrid pesticidal toxin according to the invention, is a homologous cross-region or site in the hybrid. The amino acid to the left of the crossover site is derived from one parent toxin, and the amino acid to the left of the crossover site is derived from the other parent toxin.

Mature protein: A protein that normally targets intracellular organelles and has had its transfer peptides removed.

Minimal promoter: promoter elements, particularly TATA elements, which are inactive or have greatly reduced promoter activity in the absence of upper activation. In the presence of suitable transcription factors, minimal promoters act to cause transcription.

Natural: The term refers to a gene present in the genome of an untransformed cell.

Naturally occurring: The term "naturally occurring" is used to describe a subject that can be found in nature, as distinct from that produced by man artificially. For example, naturally occurring proteins or nucleotide sequences present in organisms (including viruses) that can be isolated from natural sources and not intentionally modified by humans in the laboratory.

Nucleic acid: The term “nucleic acid” means deoxyribonucleotides or ribonucleotides or polymers thereof in single- or double-stranded form. Unless defined otherwise, the term includes nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise specified, a particular nucleic acid sequence potentially includes its conservatively modified variations (eg, degenerate codon substitutions) and complementary sequences and sequences explicitly specified. Specifically, degenerate codon substitutions can be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues. See Batzer et al. , Nucleic Acid Res. 19: 5081 (1991); Ohtsuka et al., J. Biol. Chem. 260: 2605-2608 (1985); Rossolini et al., Mol. Cell. Probes 8: 91-98 (1994). The term “nucleic acid” or “nucleic acid sequence” may be used interchangeably with genes, cDNAs, and mRNAs encoded by genes.

"ORF" means Open Reading Frame.

By “part” of a protein is meant a peptide contained in the protein and having at least 80% of its contiguous sequence.

"Plant" is any plant, especially seed plant, at any stage of development.

"Plant cells" are structural and physiological units of plants, including protoplasts and cell walls. Plant cells may be in the form of isolated single cells or cultured cells or may be part of higher organized units such as, for example, plant tissues, plant organs or complete plants.

"Plant cell culture" means a culture of plant units such as, for example, protoplasts at various stages of development, cell culture cells, cells in plant tissues, pollen, pollen tubes, embryos, knapsacks, conjugates and embryos.

"Plant" means a leaf, stem, root, flower or flower part, fruit, pollen, egg cell, conjugate, seed, cutting, cell or tissue culture or other part or product of a plant.

A "plant organ" is a visually structured and differentiated discrete part of a plant such as roots, stems, leaves, flower buds or embryos.             

As used herein, "plant tissue" refers to a group of plant cells organized into structural and functional units. Any tissue of a plant in a plant or in culture is included. The term includes, but is not limited to, any group of complete plants, plant organs, plant seeds, tissue cultures, and plant cells organized into structural and / or functional units. The use of the term in connection with or without any particular type of plant tissue as described above or included in this definition does not exclude any other type of plant tissue.

A "promoter" is an untranslated DNA sequence that contains a binding site for RNA polymerase II and is located on top of a coding region that initiates transcription of DNA. Promoter regions may also include other elements that act as regulators of gene expression.

A "protoplasm" is an isolated plant cell that contains no cell wall or contains only a portion of the cell wall.

Purification: The term "purified", when applied to a nucleic acid or protein, indicates that the nucleic acid or protein is essentially free of other cellular components involved in its natural state. It may be anhydrous or in aqueous solution but is preferably homogeneous. Purity and homogeneity are typically measured using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high performance liquid chromatography. Proteins, the major species present in the formulation, are substantially purified. The term "purified" refers to the nucleic acid or protein producing essentially one band in an electrophoretic gel. In particular, the term means that the nucleic acid or protein is at least 50% pure, more preferably at least 85%, most preferably at least about 99% pure.

Two nucleic acids are “recombined” when sequences from each of these two nucleic acids are combined as progeny nucleic acids. Both sequences are recombined "directly" if both nucleic acids are substrates for recombination. Both sequences are “indirectly” recombined when these sequences are recombined using an intermediate such as a cross oligonucleotide. In indirect recombination, only one sequence is the actual substrate for recombination, and in some cases no sequence is the substrate for recombination.

"Regulatory element" means a sequence that is involved in regulating the expression of a nucleotide sequence. Regulatory elements include promoters and termination signals operably linked to the nucleotide sequence of interest. They also typically include sequences required for proper transcription of the nucleotide sequence.

Substantially identical: In the category of two nucleic acid or protein sequences, the phrase “substantially identical” when compared and aligned for maximum correspondence, as determined by visual inspection or using one of the following sequence comparison algorithms By at least 60%, preferably at least 80%, more preferably at least 90%, even more preferably at least 95% and most preferably at least 99% nucleotide or amino acid residue identity. Preferably substantially identity is shown over a region of a sequence that is at least about 50 residues in length, more preferably a region of at least about 100 residues, and most preferably the sequence is substantially identical at least about 150 residues. In the most preferred embodiment, the sequences are substantially identical over the entire length of the coding region. In addition, substantially identical nucleic acid or protein sequences perform substantially the same function.

In sequence comparison, typically one sequence acts as a reference sequence to which the test sequence is compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, designing subsequence coordinates and designing sequence algorithm program parameters as needed. The sequence comparison algorithm then calculates the% sequence identity of the test sequence (s) relative to the reference sequence based on the designed program parameters.

Optimal sequence alignments for comparison are described in Smith & Waterman, Adv. Appl. Math. 2: 482 (1981), the local homology algorithm, Needleman & Wunsch, J. Mol. Biol. 48: 443 (1970), the homology algorithm of Pearson & Lipman, Proc. Natl'l. Acad. Sci. USA 85: 2444 (1989), a search for similarity methods, computerized implementation of the algorithms (GAP, BESTFIT, FASTA and TFASTA, manufactured by Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI). Or by visual inspection (Ausubel et al.).

One example of a suitable algorithm for determining% sequence identity and sequence similarity is Altschul et al., J. Mol. Biol. 215: 403-410 (1990). Software for performing BLAST analyzes is publicly available through the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov/). The algorithm generates a high score sequence pair (HSP) by identifying a short length word W in the query sequence that matches or satisfies some positive-evaluated threshold score T when aligned with words of equal length in the database sequence. First associated with identification (Altschul et al., 1990). This early close hit of words serves as the basis for initiating the search to find longer HSPs containing them. The word hit then expands in two directions along each sequence as long as the cumulative alignment score can be increased further. Cumulative scores are calculated using the parameters M (reward score for pairs of matched residues; always> 0) and N (penalty score for mismatched residues; always <0) for nucleotide sequences. For amino acid sequences, the scoring matrix is used to calculate the cumulative score. Expansion of the word hit in each direction results in a cumulative alignment score reduced by an amount of X from its maximum achieved value and the cumulative score approaching zero or less due to the accumulation of one or more negative-scoring residue alignments or the end of the sequence. It stops when it reaches. The BLASTN algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLAST program (for nucleotide sequences) uses a comparison of the two sequences as word length 11, expected value E 10, cutoff 100, M = 5, N = -4 as default. For amino acid sequences, the BLASTP program uses word length (W) 3, expected value (E) 10 and BLOSUM62 scoring matrices as defaults [Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89: 10915 (1989).

In addition to calculating% sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between the two sequences. See Karlin & Altschun, Proc. Nat'l. Acad. Sci. USA 90: 5873-5787 (1993). One measure of similarity provided by the BLAST algorithm is the minimum total probability (P (N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences will occur by chance. For example, a test nucleic acid sequence is considered similar to a reference sequence when the test nucleic acid sequence is compared to the reference nucleic acid sequence when the minimum total probability is less than about 0.1, more preferably less than about 0.01 and most preferably less than about 0.001. .

Another indication that two nucleic acid sequences are substantially identical is whether the two molecules hybridize to each other under stringent conditions. The phrase “hybridizing specifically” refers to binding, duplication or hybridization to only this particular nucleotide sequence under stringent conditions when the particular nucleotide sequence is present in a complex mixture (eg, of whole cells) DNA or RNA. I mean. "Substantially binding" means complementary hybridization between a probe nucleic acid and a target nucleic acid, with a small number of mismatches that can be accommodated by reducing the stringency of the hybridization medium to effect the desired detection of the target nucleic acid sequence. Include.

In the scope of nucleic acid hybridization experiments such as Southern and Northern hybridizations, "strict hybridization conditions" and "strict hybridization wash conditions" are sequence dependent and different under different environmental parameters. Longer sequences hybridize specifically at higher temperatures. For extensive guidance on hybridization of nucleic acids, see Tijssen (1993) Laboratory Techniques in Biochemistry and Molecular Biology-hybridization with Nucleic Acid Probes part I chapter 2 "Overview of principles of hybridization and the strategy of nucleic acid probe assays. "Elsevier, New York." In general, highly stringent hybridization and wash conditions are selected such that about 5 ° C. below the thermal melting point (T _m ) for a particular sequence at a given ionic strength and pH. Typically, a probe can hybridize with its target substring under “strict conditions” but not with other sequences.

T _m is the temperature at which 50% of the target sequence hybridizes to a perfectly matched probe (under some ionic strength and pH). Very stringent conditions are chosen to be equal to the T _m for a particular probe. Stringent hybridization conditions for hybridization of complementary nucleic acids with more than 100 complementary residues on a filter in Southern or Northern blots are to perform hybridization overnight at 42 ° C. with 50% formamide and 1 mg of heparin. An example of highly stringent washing conditions is 0.15 M NaCl for 15 minutes at 72 ° C. An example of stringent washing conditions is a 0.2 × SSC wash for 15 minutes at 65 ° C. (Sambrook, Infra, on the description of SSC buffer). Often, high stringency washes proceed with low stringency washes to remove background probe signals. For example, an example of medium stringency washing for duplexes of 100 or more nucleotides is 1 × SSC for 15 minutes at 45 ° C. For example, an example of a low stringency wash for duplexes of 100 or more nucleotides is 4-6 × SSC for 15 minutes at 40 ° C. For short probes (e.g., about 10 to 50 nucleotides) stringent conditions are typically at salt concentrations of Na ions of less than about 1.0 M, typically from about 0.01 to 1.0 M Na ion concentration, at pH 7.0 to 8.3 Or other salts) and the temperature is typically at least about 30 ° C. Stringent conditions may also be achieved by the addition of destabilizing agents such as formamide. In general, a signal to noise ratio that is twice (or more) than that observed in unrelated probes in a particular hybridization assay indicates detection of specific hybridization. Even nucleic acids that do not hybridize to each other under stringent conditions are still substantially identical if the proteins they encode are substantially identical. This occurs, for example, when a copy of a nucleic acid is produced using the maximum codon degeneracy allowed by the genetic code.

The following is an example of a set of hybridization / wash conditions that can be used to clone homologous nucleotide sequences that are substantially identical to the reference sequences of the present invention: The reference nucleotide sequence is preferably 7% sodium dodecyl sulfate (SDS) at 50 ° C. In 0.5 M NaPO ₄ , 1 mM EDTA and at 50 ° C., 2 × SSC, 0.1% SDS, more preferably at 50 ° C., 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO ₄ , 1 mM EDTA 1 × SSC at 50 ° C., wash in 0.1% SDS, more preferably 7% sodium dodecyl sulfate (SDS) at 50 ° C., 0.5M NaPO ₄ , 0.5 × SSC, 0.1% SDS at 50 ° C. Wash at, preferably at 7% sodium dodecyl sulfate (SDS) at 50 ° C., 0.5 M NaPO ₄ , 1 mM EDTA and at 50 ° C. at 0.1 × SSC, at 0.1% SDS, more preferably at 50 ° C. 7% Sodium Dodecyl Sulfate (SDS), 0.5 M NaPO ₄ , 0.1 × SSC, 0. at 1 mM EDTA and at 50 ° C. Hybridize under wash conditions in 1% SDS.

A further indicator that two nucleic acid sequences or proteins are substantially identical is that the protein encoded by the first nucleic acid is immunologically cross-reactive or specifically binds to the protein encoded by the second nucleic acid. Thus, if a protein is typically substantially identical to a second protein, then this is the case, for example, when the two proteins differ only in conservative substitutions.

The phrase “specifically (or selectively) binds to an antibody” or “specifically (or selectively) immunoreactive with” refers to a protein or peptide and the presence of a heterogeneous population of proteins and other biologics Under a binding reaction that is a determinant of the presence of a protein. Thus, under the designed immunoassay conditions, the specified antibody binds to a particular protein and does not bind to other proteins present in the sample in significant amounts. Specific binding to antibodies under these conditions requires an antibody selected for specificity for the particular protein. For example, an antibody generated against a protein containing an amino acid sequence encoded by any of the nucleic acid sequences of the present invention may be selected so that the immunoreactivity with other proteins except for the immunoreactivity or polymorphic variants is specific. Non-antibodies can be obtained. Various immunoassay formats can be used to select antibodies that are specifically immunoreactive with specific proteins. For example, solid phase ELISA immunoassays, Western blots or immunohistochemistry are commonly used to select immunoreactive adult monoclonal antibodies specifically for proteins. [Immune can be used to measure specific immunoreactivity. For assay format and conditions, Harlow and Lane (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, New York "Harlow and Lane"). Typically, the specific or selective response may be two or more times background signal or noise, more typically 10 to 100 times or more background signal or noise.

"Subsequence" means a sequence of nucleic acids or amino acids comprising a portion of each longer nucleic acid or amino acid (eg protein) sequence.

"Synthetic" means a nucleotide sequence that includes structural features not present in the native sequence. For example, artificial sequences that are more closely similar to the G + C content and normal codon distribution of monocotyledonous and / or dicotyledonous genes are referred to as synthetic.

"Transformation" is the process of introducing a heterologous nucleic acid into a host cell or organism. In particular, “transformation” means the stable insertion of a DNA molecule into the genome of an organism of interest. Transformed cells, tissues or insects are understood to include not only the end product of the transformation process but also the transgenic offspring thereof.

"Transformed / transformed / recombinant" means a host organism, such as a bacterium or plant, into which a heterologous nucleic acid molecule has been introduced. Nucleic acid molecules can be stably inserted into the genome of a host or can exist as extrachromosomal molecules. Such extrachromosomal molecules can autonomously replicate. Transformed cells, tissues or plants are understood to include not only the end product of the transformation process, but also their transformed progeny. By "untransformed", "untransformed" or "unrecombinant" host is meant a bacterium or plant that does not contain a wild type organism, eg, a heterologous nucleic acid molecule.

Nucleotides are specified as bases by the following standard abbreviations: adenine (A), cytosine (C), thymine (T) and guanine (G). Likewise amino acids are designated by the following standard abbreviations: alanine (Ala; A), arginine (Arg; R), asparagine (Asn; N), aspartic acid (Asp; D), cysteine (Cys; C), glutamine (Gln; Q), glutamic acid (Glu; E), glycine (Gly; G), histidine (His: H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M ), Phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y) and valine (Val; V) . In addition, (Xaa; X) represents an arbitrary amino acid.

The present invention relates to novel nucleic acid sequences that express and produce new toxins and methods of preparing and using the toxins to control pests. In particular, the present invention provides synthetic gene sequences optimized for expression in plants wherein the toxin moiety encodes various forms of hybrid Bacillus thuringiensis delta-endotoxin H04 containing domains I and II of Cry1Ab and domain III of Cry1C. It is about. Hybrid genes encoding the H04 hybrid toxin, constructed from native Cry1Ab and Cry1C genes, are described in US Pat. No. 5,736,131 and in De Maagd et al., Appl. Environ. Microbiol. 62 (5): 1537-1543 (1996). Preferred methods for constructing the synthetic H04 gene of the present invention are described in WO 93/07278. Hybrid Bacillus thuringiensis toxins encoded by this novel gene sequence are highly active against economically important pests such as chestnut moth, pink cotton seedling, tobacco moth, European light moth and cabbage moth. Hybrid Bacillus thuringiensis toxins can be used in many insect control strategies to achieve maximum efficiency with minimal impact on the environment.

The present invention includes a DNA molecule comprising a nucleotide sequence encoding a pesticidal toxin of the present invention. The present invention further includes a recombinant vector comprising the nucleic acid sequence of the present invention. In such vectors, the nucleic acid sequence is preferably contained in an expression cassette comprising regulatory elements for expressing the nucleotide sequence in a host cell capable of expressing the nucleotide sequence. Such regulatory elements typically include a promoter and a termination signal, and preferably also include elements that allow the protein encoded by the nucleic acid sequences of the invention to be efficiently transcribed. The vector comprising the nucleic acid sequence is usually used for amplifying the nucleic acid sequence of the present invention in the host cell since it can replicate particularly preferably as an extrachromosomal molecule in the host cell. In one embodiment, host cells for such vectors include bacteria, in particular Bacillus thuringiensis or E. coli. There are microorganisms such as E. coli. In other embodiments, such host cells for recombinant vectors include endogenous or complex plants. Preferred host cells for such vectors include eukaryotic cells such as plant cells. Plant cells such as corn cells are the most preferred host cells.

In a particularly preferred embodiment, the pesticidal toxins of the invention are expressed in plants. In this case, the transgenic plant expressing an effective amount of toxins protects the plant itself from pests. When insects feed on these transgenic plants, they also consume the expressed toxins. This can interfere with insects eating into plant tissue or even damage or even kill insects.

The nucleic acid sequences described herein can be incorporated into plant cells using conventional recombinant DNA techniques. In general, this involves inserting a coding sequence of the invention into an expression system that is heterologous to the coding sequence (ie, no such coding sequence exists) using standard cloning procedures known in the art. The vector contains elements essential for the transcription and translation of the inserted protein-coding sequence. Many vector systems known in the art can be used, such as plasmids, bacteriophage viruses and other modified viruses. Suitable vectors include viral vectors (e.g. lambda vector systems λgtl1, λgtl0 and Charon 4), plasmid vectors (e.g. pBI121, pBR322, pACYC177, pACYC184, pAR series, pKK223-3, pUC8, pUC9, pUC18, pUC19, pLG339, pRK290, pKC37, pKC101, pCDNAII) and other similar systems. The transformed cells can be regenerated into complete plants so that the nucleotide sequences of the invention confer insect resistance to the transgenic plants.

Plants transformed according to the present invention may be monocots or dicotyledons, corn, wheat, barley, rye, sweet potatoes, green beans, peas, chicory, lettuce, cabbage, cauliflower, broccoli, turnips, radishes, spinach, asparagus , Onion, garlic, pepper, celery, squash, pumpkin, hemp, zucchini, apple, pear, quince, melon (e.g. watermelon), plum, cherry, peach, nectarine Peach, Apricot, Strawberry, Grape, Raspberry, Blackberry, Pineapple, Avocado, Papaya, Mango, Banana, Soybean, Tomato, Sorghum, Sugarcane, Beets, Sunflowers, Rape, Clover, Tobacco, Carrots, Cotton, Alfalfa, Potatoes , Branches, cucumbers, Arabidopsis, and woody plants such as conifers and deciduous trees. Once the desired nucleotide sequence is transformed into a particular plant species, it can be propagated within that species or can be transferred to other variants of the same species, including commercially available varieties, using conventional breeding techniques.

Nucleotide sequences of the invention may require modification and optimization for expression in transgenic plants. In many cases, genes from microorganisms can be expressed at high levels in plants without modification, but low expression in transgenic plants can result from heterologous nucleotide sequences with undesirable codons in the plant. It is known in the art that all organisms exhibit particular preferences for codon usage and that the codons of the nucleotide sequences described herein can be modified to suit plant preferences while maintaining the encoded amino acids. Moreover, high expression in plants is optimally achieved from coding sequences having a GC content of at least about 35%, preferably at least about 45%, more preferably at least about 50% and most preferably at least about 60%. . Nucleotide sequences of microorganisms with low GC content may be poorly expressed in plants due to the presence of ATTTA motifs that may destabilize the message and AATAAA motifs that may cause inappropriate polyadenylation. Although preferred gene sequences may be appropriately expressed in both monocotyledonous and dicotyledonous species, as sequences appear to differ in preference (Murray et al., Nucl. Acids Res. 17: 477-498 (1989))] can be modified to be responsible for particular codon preferences and GC content preferences of monocots or dicots. Nucleotides also screen for the presence of anomalous splicing sites that can cause message cleavage. All modifications that need to be made in the nucleotide sequence as described above are gene synthesis using site directed mutagenesis, PCR, etc., and the methods described in published patent applications EP 0 385 962, EP 0 359 472 and WO 93/07278. It is accomplished using well known techniques such as.

Sequences adjacent to the starting methionine may require modification for efficient initiation of translation. For example, a sequence known to be effective in a plant can be inserted and modified. Josi has reported consensus appropriate for plants (NAR 15: 6643-6653 (1987)) and Clontech has suggested additional consensus detoxification initiators [Ref. : 1993/1994 catalog, page 210]. These consensus are suitable for use with the nucleotide sequences of the invention. The sequences are nucleotide sequences up to and including ATG (second amino acid remains unmodified) or up to and including GTC following ATG (possibly the second amino acid of the transgene is likely to be modified). It is inserted into the construct, including.

Expression of nucleotide sequences in transgenic plants is driven by promoters that have been shown to be functional in the plant. The choice of promoter can vary depending on the spatiotemporal requirements for expression and the target species. Thus, the expression of the nucleotide sequences of the invention in leaves, fruits, inflorescences (eg, inflorescences, cones, cobs, etc.), roots and / or seeds is preferred. However, in many cases, protection against more than one kind of pest is required, so expression in multiple tissues is preferred. Many promoters of dicotyledons have been shown to operate on monocots and conversely, many promoters of monocotyledonous plants have been shown to operate on dicotyledonous plants. It is ideal to be. However, there are no restrictions on the origin of the selected promoter, i.e., it is sufficient that these promoters act to drive the expression of nucleotide sequences in the cells of interest.

Preferred promoters that are structurally expressed include promoters from genes encoding actin or ubiquitin, and CaMV 35S and 19S promoters. Nucleotide sequences of the invention can also be expressed under the control of chemically regulated promoters. This can cause the insecticidal toxin to be synthesized only if the crop is treated with derived chemicals. Preferred techniques for chemical induction of gene expression are described in detail in published application EP 0 332 104 and US Pat. No. 5,614,395. A preferred promoter for chemical induction is the tobacco PR-1a promoter.

A preferred category of promoter is one that is wound inducible. Many promoters are described that express at the wound site and at the site of infection of phytopathogens. Ideally, such promoters are only active locally at the site of infection, and in this way only the pesticidal toxins needed to synthesize pesticidal toxins that kill invading pests are accumulated in the cell. Preferred promoters of this kind are described in Stanford et al., Mol. Gen. Genet. 215: 200-208 (1989), Xu et al., Plant Molec. Biol. 22: 573-588 (1993), Logemann et al., Plant Cell 1: 151-158 (1989), Rohrmeier & Lehle, Plant Molec. Biol. 22: 783-792 (1993), Firek et al. Plant Molec. Biol. 22: 129-142 (1993) and Warner et al., Plant J. 3: 191-201 (1993).

Preferred tissue specific expression patterns include green tissue specific, root specific, stem specific, fruit specific and flower specific. Suitable promoters for expression in green tissues include many that regulate genes involved in photosynthesis, many of which have been cloned from monocotyledonous and dicotyledonous plants. Preferred promoters are corn PEPC promoters from the phosphoenol carboxylase gene. See Hudspeth & Grula, Plant Molec. Biol. 12: 579-589 (1989). Preferred promoters for root specific expression are described in de Framond, FEBS 290: 103-106 (1991); EP 0 452 269, a corn metallothinein type promoter. Preferred stem specific promoters are those promoters that drive the expression of the maize trpA gene as described in US Pat. No. 5,625,136.

Particularly preferred embodiments of the invention are transgenic plants which express one or more of the nucleotide sequences of the invention in a preferred or root-specific manner in the roots. A further preferred embodiment is a transgenic plant that expresses the nucleotide sequence in a wound-induced or pathogen infection-induced manner.             

In addition to the selection of suitable promoters, constructs for expressing pesticidal toxins in plants require an appropriate transcription terminator that is linked to the bottom of the heterologous nucleotide sequence. Some of these terminators are available and are known in the art (eg tm1 from CaMV, E9 from rbcS). All available terminators known to function in plants can be used in the present invention.

Many other sequences can be incorporated into the expression cassettes for the DNA molecules described herein. These sequences include sequences that have been shown to enhance expression, such as intron sequences (eg, intron sequences from Adh1 and bronze1) and viral leader sequences (eg, leader sequences from TMV, MCMV, and AMV).

It may be desirable for the expression of the nucleotide sequences of the present invention to target different intracellular localization in plants. In some cases, localization in the cytoplasm may be desirable, while in other cases localization in some blast organelles may be desirable. Subcellular localization of the transgene encoding the enzyme is accomplished using techniques well known in the art. Typically, DNA encoding a target peptide from a gene product targeting a known organelle is engineered and fused to the top of the nucleotide sequence. Many of these target fragments are known for chloroplasts and their action in heterologous constructs is shown. Expression of the nucleotides of the invention also targets vesicles or vacuoles of the host cell. Techniques for achieving this are well known in the art.

Suitable vectors for plant transformation are also described elsewhere herein. While binary vectors with one or more T-DNA border sequences or vectors with one or more T-DNA border sequences are suitable for Agrobacterium-mediated transformation, any vector is suitable for direct gene transfer and is of interest. Linear DNA containing only existing constructs may be desirable. In the case of direct gene transfer, transformation or cotransformation into a single DNA species can be used. Schocher et al. Biotechnology 4: 1093-1096 (1986)]. For both direct gene transfer and Agrobacterium-mediated delivery, transformation is usually (but not necessarily necessary) to provide resistance to antibiotics (eg kanamycin, hygromycin or methotrexate) or herbicides (vasta). This is done using selectable markers. However, examples of such markers include neomycin phosphotransferase, hygromycin phosphotransferase, dihydrofolate reductase, phosphinothricin acetyltransferase, 2, 2-dichloropropionic acid dehalogenase, aceto Hydroxy acid synthase, 5-enolpyrubil-succimate-phosphate synthase, haloarylnitrilease, protophorhirinogen oxidase, acetyl-coenzyme A carboxylase, dihydrophteroate synthase, chloramphenicol acetyl Transferases and β-glucuronidase. Another class of markers that provides a clear choice is the mannose-6-phosphate isomerase (MPI / PMI) gene, which provides the ability to metabolize mannose mannose-6-phosphate isomerase. However, the selection of selectable or selectable markers for plant transformation is not critical to the present invention.

The recombinant DNA described above can be introduced into plant cells in a number of ways that are recognized in the art. Those skilled in the art will understand that the choice of method depends on the type of plant to be transformed. Suitable methods for transforming plant cells include microinjection (Crossway et al., BioTechniques 4: 320-334 (1986)), electroporation [Riggs at al, Proc. Natl. Acad. Sci. USA 83: 5602-5606 (1986)], Agrobacterium-mediated transformation (Hinchee et al., Biotechnology 6: 915-921 (1988); Ishida et al., Nature Biotechnology 14: 745-750 (June 1996) (relative to corn transformation), direct gene transfer [Paszkowski et al., EMBO J. 3: 2717-2722 (1984); Hayashimoto et al., Plant Physiol. 93: 857-863 (1990)] (rice), and Agratus, Inc. (Madison, Wisconsin) and Dupont Inc. (Dupont, Inc., Wilmington, Dalaware). Ballistic particle acceleration using equipment is included [Sanford et al., US Pat. No. 4,945,050; And McCabe et al., Biotechnology 6: 923-926 (1988); Weissinger et al., Annual Rev. Genet. 22: 421-477 (1988); Sanford et al., Particulate Science and Technology 5: 27-3791987) (onions); Svab et al., Proc. Natl. Acad. Sci. USA 87: 8526-8530 (1990) (tobacco chloroplasts); Christou et al., Plant Physiol. 87: 671-674 (1998) (soybeans); McCabe et al., Bio / Technology 6: 923-926 (1998) (soy); Klein et al., Proc. Natl. Acad. Sci. USA, 85: 4305-4309 (1998) (corn); Klein et al., Bio / Technology 6: 559-563 (1988) (corn); Klein et al., Plant Physiol. 91: 440-444 (1998) (corn); Fromm et al., Bio / Technology 8: 833-839 (1990); and Gordon-Kamm et al., Plant Cell 2: 603-618 (1990) (corn); Koziel et al., Biotechnology 11: 194-200 (1993) (corn); Shimamoto et al., Nature 338: 274-277 (1998) (rice); Christou et al., Biotechnology 9: 957-962 (1991) (rice); Datta et al., Bio / Technology 8: 736-740 (1990) (rice); European Patent Application EP 0 332 581 (duck and other rice family); Vasil et al., Biotechnology 11: 1153-1558 (1993) (wheat); Weeks et al., Plant Physiol. 102: 1077-1084 (1993) (wheat); Wan et al., Plant physiol. 104: 37-48 (1994) (barley); Jahne et al., Theor. Appl. Genet. 89: 525-533 (1994) (barley); Umbeck et al., Bio / Techonology 5: 263-266 (1987) (cotton); Casas et al., Proc. Natl. Acad. Sci. USA 90: 11212-11216 (Dec. 1993) (sorghum); Somer et al., Bio / Technology 10: 1589-1594 (Dec. 1992) (oats), Torbert et al., Plant Cell Reports 14: 635-640 (1995) (oats); Weeks et al., Plant Physiol. 102: 1077-1084 (1993) (wheat); Chang et al., WO 94/13822 (wheat) and Nehra et al., The Plant Journal 5: 285-297 (1994) (wheat)]. Particularly preferred embodiments for introducing recombinant DNA molecules into maize by microprojectile bombardment are described in Koziel et al., Biotechnology 11: 194-200 (1993), Hill et al., Euphytica 85 : 119-123 (1995) and Koziel et al., Annals of the New York Academy of Sciences 792: 164-171 (1996). A further preferred embodiment is the protoplast transformation method for corn, described in EP 0 292 435. Plant transformation may consist of a single DNA species or multiple DNA species (ie, cotransformation) and these two techniques are suitable for use with the coding sequences of the present invention.

In another preferred embodiment, the nucleotide sequences of the invention are transformed directly into the plasmid genome. The main advantage of chromatin transformation is that the pigment can generally express bacterial genes without sufficient modification and can express multiple open reading frames under the control of a single promoter. Chromosome transformation techniques are described in US Pat. Nos. 5,451,513, 5,545,817 and 5,545,818, PCT Application WO 95/16783, and McBride et al. (1994) Proc. Natl. Acad. Sci. USA 91, 7301-7305. Basic techniques for chloroplast transformation include genes of interest with regions of cloned chromatin DNA flanked in selectable markers, for example using biologic or protoplast transformation (eg calcium chloride or PEG mediated transformation). Is introduced into a suitable target tissue. The 1-1.5 kb flanking region designated as the target sequence facilitates homologous recombination with the chromosomal genome, thereby allowing replacement or modification of specific regions of the plasma. Initially, the point mutation in chloroplast 16S rRNA and the rps12 gene, which provides resistance to spectinomycin and / or streptomycin, are used as selectable markers for transformation. Svab, Z., Hajdukiewicz, P ., and Maliga, P. (1990) Proc. Natl. Acad. Sci. USA 87, 8526-8530; Staub, J. M., and Maliga, P. (1992) Plant Cell 4, 39-45]. This results in a stable monocyclic transformant at about 1 frequency per 100 collisions of the target leaf. The presence of a cloning site between these markers allowed the formation of a chromatin target vector for introduction of foreign genes (Staub, J. M., and Maliga, P. (1993) EMBO J. 12: 601-606). Substantial increase in the frequency of transformation is obtained by replacing the recessive rRNA or r-protein antibiotic resistance gene with a dominant selectable marker (the bacterial aadA gene encoding spectinomycin-detoxifying enzyme aminoglycoside-3'-adenyltransferase) [Svab, Z., and Maliga, P. (1993) Proc. Natl. Acad. Sci. USA 90, 913-917. Previously, this marker has been used successfully for high-frequency transformation of the chromatin genome of the green alga Chlamydomonas Reinhardty (Goldschmidt-Clermont, M. (1991) Nucl. Acids Res. 19, 4083-4089. Other selectable markers useful for chromatin transformation are known in the art and are included within the scope of the present invention. Typically, about 15 to 20 cell division cycles after transformation are required to reach a monomorphic state. Chromosome expression, in which genes are inserted into all thousands of copies of the cyclic chromatin genome present in each plant cell by homologous recombination, is a nuclear-expressed gene that allows expression levels that can easily exceed 10% of the total soluble plant protein It uses a huge number of copies more advantageous than. In a preferred embodiment, the nucleotide sequence of the present invention is inserted into a pigment target vector and transformed into the pigment genome of the desired plant host. A plant is obtained that is monomorphic with respect to the chromatin genome containing the nucleotide sequence of the present invention, and is preferably capable of highly expressing the nucleotide sequence.

The invention will be further explained with reference to the following detailed examples. These embodiments are provided for illustration only and are not limited unless specified otherwise. Standard recombinant DNA and molecular cloning techniques used herein are well known in the art and described in Ausubel (ed.), Current Protocols in Molecular Biology, John Wiley and Sons, Inc. (1994); T. Maniatis, EF Fritsch and J. Sambrook, Molecular Cloning: A Laboratory manual, Cold Spring Harbor laboratory, Cold Spring Harbor, NY (1989); And TJ Silhavy, MLBerman and LW Enquist, Experiments with Gene Fusions, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1984).

Example 1: Expression and Purification of H04 Toxin Fragment

A form cleaved from the H04 hybrid toxin gene encoding essentially the Bt toxin consisting of domains I and II of Cry1Ab and domain III of Cry1C (De Maagd et al., Appl. Environ. Microbiol. 62 (5) : 1537-1543 (1996). Cloned into expression vectors such as pBluescript SK-, Bacillus shuttle vector or pET 21b (+) for overexpression in E. coli. Cells are grown on a 37 ° C. shaker (250 rpm) for 24 to 48 hours in LB medium containing 50 μg / ml ampicillin. Cells are harvested by centrifugation at 7,000 rpm for 10 minutes. The pellet is sonicated with a 2 second pulse for 10 minutes using a Bronson sonicator. Complete sonication is checked under a microscope. Soluble fractions are removed by centrifugation at 10,000 rpm for 10 minutes. The resulting pellet containing crystalline protein is washed 4-5 times with 2% Triton X-100 containing 0.5M NaCl. Wash sequentially with 0.5 M NaCl (4-5 times) and wash the final pellet with distilled water (2 times). The resulting pellets are soluble in 50 mM Na ₂ CO ₃ buffer containing 10 mM dithiothreitol at 37 ° C. for 2 hours. Solubilized protein is separated from insoluble material by centrifugation at 12,000 rpm for 10 minutes. Protein samples are dialyzed with 50 mM Na ₂ CO ₃ buffer (pH 9.0) for bioassay.

Example 2: Bioassay

LC50 is performed in chestnut moth, pink cotton worm, tobacco moth and European light moth using, for example, purified truncated H04 protein produced as described above in Example 1. The result is:

LC50 Night Moth 133ng / cm ²

LC50 Pink Cotton Seed Bug 691 ng / cm ²

LC50 Tobacco Moth 299ng / cm ²

LC50 European Lighting Moth 31 ng / cm ²

Example 3: Synthetic H04 Gene Construction

Synthetic nucleotide sequences encoding the toxic moiety of H04 can be found in the corn preference codon table [Murray et al., Nucl Acids Res. 17: 477-498, 1989; It is incorporated herein by reference, using the "Backtranslation" program developed by the University of Wisconsin GCG Program Group, which is described in De Maagd et al., Appl. Environ. Microbiol. 62 (5): 1537-1543 (1996)] is designed by backtrnaslating the amino acid sequence of the H04 hybrid toxin fragment (domains I and II of Cry1Ab and domain III of Cry1C). Preferably, the most frequently used corn codon is used for each amino acid as described in WO 93/07278.

Synthetic nucleotide sequences encoding the toxin portion of H04 can be constructed as several fragments. Each fragment is constructed by hybridizing 10 pairs of oligomers of 60 to 75 nucleotides in length that represent both chains of the gene. About 15 nucleotide overlaps are designed between sequential oligonucleotide pairs for accurate orientation and aggregation. Oligonucleotides can be constructed, for example, by Genosys Biotechnologies Inc., TX. Each pair of oligomers is hybridized and phosphorylated using, for example, enzyme polynucleotide kinase (New England Biolabs, Inc., MA) under conditions specified by the salesman. Phosphorylated fragment pairs are then hybridized and linked into a high replicate plasmid vector containing, for example, an ampicillin resistance gene, and transformed, eg, into soluble DH5α cells. The cells are plated in ampicillin containing medium and incubated overnight at 37 ° C. Colonies are selected for inserted DNA. Sequencing the DNA and selecting clones containing the correct sequence. The fragments are then linked by restriction digestion, ligation and transformation using unique restriction sites between the fragments.                 

SEQ ID NO: 3 shows a synthetic nucleotide sequence that encodes the 631 amino acid toxin site of H04 (contains no pretoxin tail region), and SEQ ID NO: 4 shows the amino acid sequence of H04 toxin encoded by the synthetic nucleotide sequence described in SEQ ID NO: 3 Indicates. SEQ ID NO: 11 shows the amino acid sequence of construct pNOV1308 containing a constitutive corn ubiquitin promoter operably linked to the synthetic H04 gene sequence set forth in SEQ ID NO: 3.

In addition to the synthetic genes described above that encode only the toxin portions of the H04 hybrid (domains I and II of Cry1Ab and domain III of Cry1C) (SEQ ID NO: 3), additional synthetic H04 genes are described in US Pat. No. 5,625,136, which is incorporated herein by reference. All or a portion of the synthetic cry1Ab tail region described in VII) is fused to the 3 'end of the H04 toxin portion.

SEQ ID NO: 5 shows a synthetic nucleotide sequence encoding a toxin portion of H04 and a synthetic nucleotide sequence encoding a full length Cry1Ab tail portion, and SEQ ID NO: 6 is the amino acid sequence of the H04 + Cry1Ab tail encoded by the synthetic nucleotide sequence described in SEQ ID NO: 5 To present. SEQ ID NO: 12 shows the nucleotide sequence of construct pNOV1436, which contains a preferred corn MTL promoter for roots, operably linked to the synthetic H04 gene sequence set forth in SEQ ID NO: 5. SEQ ID NO: 13 shows the nucleotide sequence of construct pNOV1441, containing a constitutive corn ubiquitin promoter operably linked to the synthetic H04 gene sequence set forth in SEQ ID NO: 5.

SEQ ID NO: 7 shows another synthetic nucleotide sequence encoding the toxin portion of the H04 and full length Cry1Ab tail portion, and SEQ ID NO: 8 shows the amino acid sequence of the H04 + Cry1Ab tail encoded by the synthetic nucleotide sequence described in SEQ ID NO: 7. SEQ ID NO: 14 sets forth the nucleotide sequence of construct pNOV1305 containing a constitutive corn ubiquitin promoter operably linked to a synthetic H04 gene sequence set forth in SEQ ID NO: 7. SEQ ID NO: 15 shows the nucleotide sequence of construct pNOV1313 containing a constitutive corn ubiquitin promoter operably linked to a synthetic H04 gene sequence set forth in SEQ ID NO: 7.

SEQ ID NO: 9 shows a synthetic nucleotide sequence that encodes the toxin portion of H04 and only the first 40 amino acids of the Cry1Ab tail, and SEQ ID NO: 10 is a Cry1Ab truncated to H04 + 40 amino acids encoded by the synthetic nucleotide sequence described in SEQ ID NO: 9 The amino acid sequence of the tail is shown. SEQ ID NO: 16 shows the nucleotide sequence of construct pNOV1435 containing a maize MTL promoter preferred for roots operably linked to the synthetic H04 gene sequence set forth in SEQ ID NO: 9. SEQ ID NO: 17 shows the nucleotide sequence of construct pZU578, containing an arabidopsis actin-2 promoter operably linked in addition to the synthetic H04 gene sequence set forth in SEQ ID NO: 9.

Example 4: Modification of Coding Sequence and Contiguous Sequence

The nucleotide sequences described herein can be modified for expression in transgenic host cells. Host plants that express the nucleotide sequence and produce insecticidal toxins in the cell exhibit enhanced resistance to insect attacks, thereby better equipping them for preventing crop losses associated with such attacks.

In plant transgenic expression of genes derived from microbial sources may require modification of the genes to achieve and optimize the gene expression in the plant. In particular, bacterial ORFs encoding individual enzymes but encoded by the same transcript in natural microorganisms are prominently expressed as individual transcripts in plants. To accomplish this, each microbial ORF is isolated separately and cloned in a cassette providing a plant promoter sequence at the 5 'end of the ORF and a plant transcription terminator at the 3' end of the ORF. An isolated ORF sequence preferably comprises an initiating ATG codon and a terminating STOP codon, but may comprise additional sequences other than the initiating ATG and STOP codons. In addition, ORFs may be cleaved but still retain the required activity, and particularly long ORFs, which are truncated forms that retain activity, may be desirable for expression in the transgenic organism. "Plant promoter" and "plant transcription terminator" refer to promoters and transcription terminators that operate in plant cells. This includes promoters and transcription terminators that can be derived from non-plant sources such as viruses (eg, cauliflower mosaic virus).

In some cases, modifications of ORF coding sequences and contiguous sequences are required. The modification is sufficient only to isolate the fragment containing the ORF of interest and insert it into the bottom of the plant promoter. For example, Gaffney et al. (Science 261: 754-756 (1993)) described the Pseudomonas nahG gene as the Pseudomonas gene x bp and STOP codon, which is located on top of ATG without altering the coding sequence. The underlying y bp has been successfully expressed in transgenic plants under the control of the CaMV 35S promoter and CaMV tml terminator while linked to nahG ORF. Preferably, as little contiguous microbial sequence as possible should be linked to the top of the ATG and the bottom of the STOP codon. In fact, such construction may depend on the availability of the restriction site.

In other cases, expression of genes derived from microbial sources can provide a problem with expression. These problems are well described in the art, and are common in genes derived from certain sources, particularly Bacillus. These problems can be applied to the nucleotide sequences of the present invention and modifications of these genes can be accomplished using techniques well known in the art. The problems you may face include:

1. Codon Usage

Preferred codon usage in plants differs from the preferred codon usage in certain microorganisms. By comparing codon usage in cloned microbial ORFs and usage in plant genes (and in particular genes from target plants), codons can preferably be identified in the ORF that should be modified. Typically, plant evolution tends to strongly favor nucleotides C and G at the third base position of the monocotyledonous plant, while dicotyledons often use nucleotides A or T at these positions. By modifying the gene to include the desired codon usage for specific target transgenic plants, many problems with GC / AT content and anomalous splicing described below can be addressed.                 

2. GC / AT content

Plant genes typically have a GC content of at least 35%. ORF sequences rich in A and T nucleotides can cause some problems in plants. First, the ATTTA motif is thought to cause message destabilization and is found at the 3 'end of many mRNAs with short half-lives. Second, the generation of polyadenylation signals such as AATAAA at inappropriate locations in the message is thought to cause premature interruption of transcription. Monocots can also recognize AT-rich sequences as splicing sites (see below).

3. Sequences Adjacent to Initiating Methionine

Plants are distinguished from microorganisms in that their messages do not have defined ribosomal binding sites. Rather, it is believed that ribosomes bind to the 5 'end of the message to search for the first valid ATG to initiate transcription. Nevertheless, there is a preference for certain nucleotides adjacent to ATG and the expression of microbial genes can be enhanced by including eukaryotic consensus transcription initiators in ATG. Clontech [Ref. 1993/1994 catalog, page 210; Which is incorporated herein by reference. It has been suggested as a consensus transcription initiator for expressing the E. coli uidA gene. In addition, Josh (NAR 15: 6643-6653 (1987), which is incorporated herein by reference), compared a number of plant sequences adjacent to ATG and presented different consensus sequences. In situations facing difficulties in expressing the microbial ORF in plants, one of these sequences may be included in the starting ATG to improve transcription. In such cases, the last three nucleotides of the consensus may not be suitable for inclusion in the modified sequence due to the modification of its second AA residue. Preferred sequences adjacent to the starting methionine may differ between different plant species. A survey of 14 corn genes in the GenBank database gives the following results:

Position Before Initiation ATG in 14 Corn Genes

The analysis can be performed on the desired plant species into which the nucleotide sequence is incorporated, and the sequence adjacent to the ATG modified to include the desired nucleotide.

4. Removal of anomalous splicing sites

Genes cloned from non-plant sources and not optimized for expression in plants may contain motifs that can produce truncated or deleted messages because they can be recognized and cleaved as 5 'or 3' splicing sites in plants. It may be. These sites can be removed using techniques well known in the art.

Techniques for modifying coding sequences and contiguous sequences are well known in the art. If it is deemed that initial expression of the microbial ORF is poor and alterations to the sequences described above are desired, the construction of synthetic genes can be accomplished according to methods well known in the art. These methods are described, for example, in Publications EP 0 385 962, EP 0 359 472 and WO 93/07278, all of which are incorporated herein by reference. In most cases, expression of the gene construct is preferably assayed using a transient assay protocol (which is well known in the art) prior to delivering the gene to the transgenic plant.

Example 5: Construction of Plant Expression Cassettes

First, the coding sequence to be expressed in the transgenic plant is collected as an expression cassette behind a suitable promoter expressible in the plant. The expression cassette may comprise any additional genes required or selected for expression of the transgene. Such sequences include, but are not limited to, transcriptional terminators, exogenous sequences that enhance expression (eg, introns), pivotal sequences, and sequences for targeting gene products to specific organ and cell compartments. This expression cassette can thus be readily delivered to the transformation vectors described below. The following is a description of the various components of a conventional expression cassette.

1. Promoter

The choice of promoter used in the expression cassette can determine the spatiotemporal expression pattern of the transgene in the transgenic plant. Selected promoters can express specific transgenes in specific cell types (e.g. leaf epithelial cells, foliar cells, root cortical cells) or in specific tissues or organs (e.g. roots, leaves or flowers) and selection of gene products The desired accumulation position can be reflected. Alternatively, the selected promoter can induce the expression of a gene under inducible conditions. Promoters vary in their strength, the ability to promote transcription. Depending on the host cell system used, any of a number of suitable promoters can be used, including the natural promoter of the gene. The following are non-limiting examples of promoters that can be used in expression cassettes.

a. Constitutive expression, ubiquitin promoters:

Ubiquitin is a gene product known to accumulate in many cell types, and its promoters have been cloned from several species for use in transgenic plants (e.g. sunflowers: Binet et al. Plant Science 79: 87- 94 (1991)], corn (Christensen et al. Plant Molec. Biol. 12: 619-632 (1989)). Corn ubiquitin promoters were developed in transgenic monocots and their sequences and vectors constructed for monocot plant transformation are described in patent publication EP 0 342 926, which is incorporated herein by reference. See also Taylor et al., Plant Cell Rep. 12: 491-495 (1993) describe a vector that contains a corn ubiquitin promoter and a first intron and is highly active in cell suspensions of a number of monocotyledonous plants when introduced via vectors and microinjection bombardment. The Arabidopsis ubiquitin promoter is ideal for use with the nucleotide sequences of the present invention. The ubiquitin promoter is suitable for gene expression in both transgenic plants, monocotyledonous and dicotyledonous plants. Suitable conformation vectors are any transformation vectors described herein, as modified by introducing a derivative of pAHC25 or an appropriate ubiquitin promoter and / or intron sequence.

b. Constitutive expression, CaMV 35S promoter:

The construction of plasmid pCGN1761 is described in published patent application EP 0 392 225 (Example 23), which is incorporated herein by reference. pCGN1761 contains a "double" CaMV 35S promoter, a tml transcription terminator and a unique EcoRI site between the promoter and terminator and has a pUC backbone. Derivatives of pCGN1761 are constructed to have modified polylinkers comprising NotI and XhoI sites in addition to existing EcoRI sites. The derivative is designated pCGN1761ENX. pCGN1761ENX is useful for cloning cDNA sequences or coding sequences (including microbial ORF sequences) in its polylinkers for expression under the control of a 35S promoter in transgenic plants. The entire 35S promoter-coding sequence-tml terminator cassette of this construct can be cleaved at the HindIII, SphI, SalI and XbaI sites located at 5 'of the promoter for delivery to a transformation vector as described below, Cleavage can be made at the XbaI, BamHI and BglI sites located 3 'of the terminator. In addition, to replace with other promoters, the double 35S promoter fragment can be removed by 5 ′ cleavage with HindIII, SphI, SalI, XbaI or PstI and 3 ′ cleavage of any site of polylinker restriction sites (EcoRI, NotI or XhoI). . If desired, modifications around the cloning site can be achieved by introducing sequences that can enhance transcription. This is particularly useful when overexpression is desired. For example, pCGN1761ENX can be modified by optimizing the transcription initiation site described in Example 37 of US Pat. No. 5,639,949, which is incorporated herein by reference.

c. Constitutive expression, actin promoter

Several isoforms of actin are known to be expressed in most cell types, so actin promoters are frequently used as structural promoters. In particular, promoters from the Act1 gene of rice have been cloned and characterized. McElroy et al. Plant Cell 2: 163-171 (1990). The 1.3 kb fragment of this promoter was found to contain all the regulatory factors necessary for expression in the protoplasts of rice. In addition, various expression vectors based on the Act1 promoter have been constructed, particularly for use in monocots. McElroy et al., Mol. Gen. Genet. 231: 150-160 (1991). These vectors include Actl-intron 1, Adh1 5 'flanking sequences and sequences from Adh1-intron 1 (from corn alcohol dehydrogenase gene) and CaMV 35S promoters. The vector with the highest expression rate is either a fusion of 35S with Act1 intron or a fusion of Act1 5 'flanking sequence with Act1 intron. Optimization of the sequence around the starting ATG (ATG of the GUS reporter gene) also enhances expression. Promoter expression cassettes described by McElroy et al. (Mol. Gen. Genet. 231: 150-160 (1991)) can be readily modified for expression of genes and are particularly well suited for use in monocotyledonous hosts. Do. For example, removing promoter-containing fragments from the McElroy construct and replacing the double 35S promoter in pCGN1761ENX can be used for specific gene sequences or insertions. The fusion gene thus constructed can be delivered to a suitable transformation vector. In a separate report, the Act1 promoter of rice with a first intron was found to induce high expression in cultured barley cells. See Chibbar et al., Plant Cell Rep. 12: 506-509 (1993).

d. Inducible Expression, PR-1 Promoter:

The dual 35S promoter in pCGN1761ENX can be replaced with any other promoter selected that can result in a suitably high expression level. For example, one of the chemically controllable promoters described in US Pat. No. 5,614,395, such as the tobacco PR-1a promoter, may replace the dual 35S promoter. Alternatively, the Arabidopsis PR-1 promoter described in Lebel et al., Plant J. 16: 223-233 (1998) can be used. The selected promoter is preferably cleaved from its source using restriction enzymes, but can be PCR-amplified using primers with appropriate terminal restriction sites. If PCR amplification is to be performed, the promoter should be resequenced after cloning the amplified promoter in the target vector to check for amplification errors. The chemically / pathogen control tobacco PR-1a promoter is cleaved from plasmid pCIB1004 (for construction, see Example 21 of EP 0 332 104, incorporated herein by reference) and transferred to plasmid pCGN1716ENX (Uknes et al. , Plant Cell 4: 645-656 (1992). pCIB1004 is cleaved with NcoI and blunt ended by treatment of the 3 ′ overhang of the linearized fragments obtained with T4 DNA polymerase. The fragment is then cleaved with HindIII and the resulting PR-1a promoter containing fragment is gel purified and cloned into pCGN1761ENX, of which 35S promoter has been removed. This is done by cleaving with XhoI, blunt ended with T4 polymerase, then cleaving with HindIII and separating the larger vector-terminator containing fragments from which the pCIB1004 promoter fragment was cloned. This gives PRpCGN1761ENX containing the PR-1a promoter and tml terminator, and an intervening polylinker containing EcoRI and NotI sites. The selected coding sequence can be inserted into the vector and then the fusion product (ie, promoter-gene-terminator) can be delivered to any selected transformation vector comprising the vector described above. Various chemical modulators, including the benzothiadiazole, isnicotinic acid and salicylic acid compounds described in US Pat. Nos. 5,523,311 and 5,614,395, can be used to induce the expression of coding sequences selected in plants transformed according to the present invention. have.

e. Inducible Expression, Ethanol-Induced Promoters:

Promoters that can be induced by certain alcohols or ketones, such as ethanol, can also be used to drive expression of the coding sequences of the invention. Such promoters include the alcA gene promoter from Aspergillus nidulans (Caddick et al. (1998) Nat Biotechnol 16: 177-180). In A. nidulans, the alcA gene encodes alcohol dehydrogenase I, whose expression is regulated by AlcR transcription factors in the presence of chemical inducers. For the purposes of the present invention, a CAT coding sequence in plasmid palcA: CAT comprising alcA gene promoter sequence fused to at least 35S promoter [Caddick et al. (1988) Nat Biotechnol 16: 177-180] is replaced with the coding sequence of the present invention to form an expression cassette with the coding sequence under the control of the alcA gene promoter. This is done using methods known in the art.

f. Inducible expression, glucocorticoid-inducible promoter

Induction of expression of nucleic acid sequences of the present invention using systems based on steroid hormones is also included in the present invention. For example, using a glucocorticoid-mediated induction system (Aoyama and Chua (1997) The Plant Journal 11: 605-612). Glucocorticoids, such as synthetic glucocorticoids, preferably dexamethasone, are preferably applied at concentrations ranging from 0.1 to 1 mM, more preferably at concentrations ranging from 10 mM to 100 mM. For the purposes of the present invention, the luciferase gene sequence is replaced with the nucleic acid sequence of the present invention to produce an expression cassette in which the nucleic acid sequence of the present invention is fused to a 35S minimal promoter under the control of six copies of the GAL4 upstream activating sequence. do. This is done using methods well known in the art. The trans-acting factor is the transactivation domain of the herpes virus protein VP16 fused to the hormone-binding domain of the rat glucocorticoid receptor (Picard et al. (1988) Cell 54: 1073-1080) [TriZenberg et al. . (1988) Genes Devel. 2: 718-729] and GAL4 DNA-binding domains [Keegan et al. (1986) Science 231: 699-704. Expression of the fusion protein is controlled by any promoter suitable for expression in plants known in the art or described herein. The expression cassette is also included in a plant comprising the nucleic acid sequence of the invention fused to a 6xGAL4 / minimal promoter. Thus, tissue-specific or organ-specificity of the fusion protein is achieved by inducing inducible tissue-specific or organ-specificity of the pesticidal toxin.

g. Root Specific Expression:

Another pattern of gene expression is root expression. Suitable root promoters are promoters of the maize metallothionein (MTL) gene described in de Framond, FEBS 290: 103-106 (1991), and US Pat. No. 5,466,785, which is incorporated herein by reference. The "MTL" promoter is transferred to a suitable vector, such as pCGN1761ENX for insertion of the selected gene, followed by transfer of the entire promoter-gene-terminator cassette to the transformation vector of interest.

h. Wound-induced promoters:

Wound-induced promoters may also be suitable for gene expression. Many such promoters are described in Xu et al. Plant Molec. Biol. 22: 573-588 (1993), Logemann et al. Plant Cell 1: 151-158 (1989), Rohrmeier & Lehle, Plant Molec. Biol. 22: 783-792 (1993), Firek et al. Plant Molec. Biol. 22: 129-142 (1993), Warner et al. Plant J. 3: 191-201 (1993), all of which are suitable for use in the present invention. Logemann et al. Describe the 5 'upstream sequence of the dicot potato wunI gene. Xu et al suggest that the wound inducible promoter (pin2) from dicotyledonous potatoes is active in monocotyledonous rice. In addition, Rohrmeier and Lehle describe the cloning of maize Wip1 cDNA which is wound-inducible and can be used to isolate cognate promoters by standard techniques. Similarly, Firek et al. Warner et al. Described wound inducible genes from monocotyledonous Asparagus officinalis expressed at local wounds and pathogen invasion sites. These promoters can be used to express the genetic wells at the plant wound site by transferring them into suitable vectors using the cloning techniques known in the art and fusion to the genes according to the invention.

i. Preferred Expressions for Carpenters:

Patent Application WO 93/07278, which is incorporated herein by reference, describes the isolation of maize trpA gene that is preferentially expressed in carpenter cells. There are gene sequences and promoters that correspond from the start of transcription to about 1726 bp. Using standard molecular biology techniques, the promoter or portion thereof can be delivered to a vector such as pCGN1761, where the promoter can be used to replace the 35S promoter to express foreign genes in a carpenter's preferred manner. Indeed, fragments containing a promoter or part thereof that is preferred for a carpenter can be delivered to any vector and usefully modified for transgenic plants.

j. Leaf-specific Expression:

Corn genes encoding phosphoenol carboxylase (PEPC) are described in Hudspeth & Grula, Plant Molec Biol 12: 579-589 (1989). Using standard molecular biology techniques, promoters of these genes can be used to drive the expression of any gene in a transgenic plant in a leaf specific manner.

k. Pollen-specific expression:                 

WO 93/07278 describes the isolation of the calcium dependent protein kinase (CDPK) gene of maize expressed in pollen cells. The gene sequence and the promoter correspond to up to 1400bp from the start of transcription. Using standard molecular biology techniques, the promoter or portion thereof can be delivered to a vector such as pCGN1761, where the promoter can be used to replace the 35S promoter to express the nucleic acid sequence of the invention in a pollen specific manner.

1. Receptor mediated transactivation in the presence of chemical ligands:

U.S. Patent 5,880,333, which is incorporated herein by reference, discloses that type II hormone receptors such as ecdysone receptor (EcR) and Ultraspiracle (USP), which act together as heterodimers, are suitable chemical ligands in plant cells, e.g. For example, systems have been described that modulate the expression of a target polypeptide in the presence of tebufenozide.

2. Warrior Terminator

Various transcription terminators can be used for the expression cassette. These transcription terminators are involved in transcription termination and precise polyadenylation following the transgene. Suitable transcription terminators are terminators known to function in plants and include CaMV 35S terminators, tml terminators, nopaline synthase terminators and pea rbcS E9 terminators. These terminators can be used for both monocotyledonous and dicotyledonous plants. In addition, a natural transcription terminator of a gene can be used.

3. Sequences for Enhancing or Regulating Expression

A number of sequences have been found to enhance gene expression in transcription units and these sequences can be used with the genes of the invention to increase expression in transgenic plants.

Various intron sequences have been shown to enhance expression, particularly in monocotyledonous cells. For example, introns of maize Adh1 gene have been shown to significantly enhance the expression of wild-type genes under cognate promoters when introduced into maize cells. Intron 1 has been shown to particularly enhance the expression of fusion constructs containing the chloramphenicol acetyltransferase gene (Callis et al., Genes Develop 1: 1183-1200 (1987)). In the same experimental system, introns from the corn bronze 1 gene also have a similar effect on enhancing expression. Intron sequences are typically contained within plant transformation vectors, usually within a non-toxic leader.

Many non-ready leader sequences derived from viruses are known to enhance expression, and such leader sequences are particularly effective in dicotyledonous cells. Specifically, leader sequences from tobacco mosaic virus (TMV, "W-sequence"), maize half lobe virus (MCMV) and alpha wave mosaic virus (AMV) have been shown to be effective in increasing expression. Gallie et al. . Nucl. Acids Res. 15: 8693-8711 (1987); Skuzeski et al. Plant Molec. Biol. 15: 65-79 (1990).

4. Targeting Gene Products Into Cells

Various mechanisms for targeting gene products are known to exist in plants, and the sequences regulating the function of these mechanisms are also described in some detail. For example, targeting of gene products to pigments (eg chloroplasts) is regulated by signal sequences found at the amino-terminal ends of various proteins and cleaved during chloroplast influx to produce mature proteins. Comai et al. . J. Biol. Chem. 263: 15104-15109 (1988). These signal sequences may be effective for fusion to heterologous gene products to introduce heterologous products into chloroplasts (van den Broeck et al., Nature 313: 358-363 (1985)). DNA encoding the appropriate signal sequence can be isolated from the 5 'end of the cDNA encoding RUBISCO protein, CAB protein, EPSP synthase enzyme, GS2 protein, and many other proteins known to be located in the chloroplast. Paragraph entitled "Expression With Chloroplast Targeting" in the examples of patent 5,639,949.

Other gene products are localized in other organelles such as mitochondria and peroxysomes. See Unger et al., Plant Molec. Biol. 13: 411-418 (1989). CDNAs encoding such products can also be engineered to target heterologous gene products to the organelles. Examples of such sequences are nuclear-encoded ATPase and aspartate amino transferase isotypes specific for mitochondria. Targeting of cellular protein bodies is described in Rogers et al. Proc.Natl.Acad.Sci. USA 82: 6512-6516 (1985).

Also described are sequences that can target gene products to other cell compartments. The amino terminal sequence is associated with targeting to ER, apoplasts and extracellular secretion from eruption cells (Kohehler & Ho, Plant Cell 2: 769-783 (1990)). In addition, the amino terminal sequence is associated with targeting of the gene product to vacuoles along with the carboxy terminal sequence. Shinshi et al. Plant Molec. Biol. 14: 357-368 (1990).

The appropriate targeting sequence described above can be fused to the transgene to direct the transgene product to any organelle or cell compartment. For chloroplast targeting, for example, chloroplast signal sequences from the RUBISCO gene, CAB gene, EPSP synthase gene or GS2 gene are fused to the amino terminal ATG of the transgene in frame. The signal sequence selected should comprise a known cleavage site and the constructed fusion should take into account any amino acids following the cleavage site necessary for cleavage. In some cases this requirement can be fulfilled by adding a few amino acids between the cleavage site and the transgene ATG, or by replacing some amino acids in the transgene sequence. Fusions constructed for chloroplast influx were then read in vitro by transcribed constructs in vitro, followed by Bartlett et al. In: Edelmann et al. (Eds.) Methods in Chloroplast Molecular Biology, Elsevier, pp 1081-1091 (1982); Wasmann et al. Mol. Gen. Genet. 205: 446-453 (1986) can be used to test for chloroplast uptake efficacy by absorbing chloroplasts in vitro. Such construction techniques are well known in the art and are equally applicable to mitochondria and peroxysomes.

The intracellular targeting mechanisms described above are not only used with cognate promoters to achieve localization targets to specific cells under transcriptional control of promoters with expression patterns different from those of the promoter from which the target signal is derived, It can also be used with heterologous promoters.

Example 6: Construction of Plant Transformation Vectors

Many transformation vectors useful for plant transformation are known to those skilled in the field of plant transformation, and genes suitable for the present invention can be used with any of these vectors. The choice of vector may depend on the desired transformation technique and the target species for transformation. For certain target species different antibiotic or pesticide selection markers may be preferred. Selection markers commonly used for transformation include the nptII gene conferring resistance to kanamycin and related antibiotics (Messing & Vieira, Gene 19: 259-268 (1982); Bevan et al., Nature 304: 184-187 (1983)], the bar gene conferring resistance to the herbicide phosphinothricin (White et al., Nucl. Acids Res 18: 1062 (1990), Spencer et al. Theor. Appl. Genet 79: 625-631 (1990)], hph gene conferring resistance to the antibiotic hygromycin (Blochinger & Diggelmann, Mol Cell Biol 4: 2929-2931) and conferring resistance to metatrexate dhfr gene (Bourouis and Jarry, EMBO J.2 (7): 1099-1104 (1983)), EPSPS gene conferring resistance to glyphosate (US Pat. Nos. 4,940,935 and 5,188,642) And mannose-6-phosphate isomerase genes (US Pat. Nos. 5,767,378 and 5,994,629) that provide the ability to metabolize mannose.                 

1. Vectors Suitable for Agrobacterium Transformation

Many vectors are useful for transformation with Agrobacterium tumefaciens. These vectors typically have one or more T-DNA border sequences and include vectors such as pBIN18 (Bevan, Nucl. Acids Res. (1984)) and pXYZ. In the following two conventional vectors suitable for Agrobacterium transformation are described.

a. pCIB200 and pCIB2001:

Dual vectors pCIB200 and pCIB2001 are used to construct recombinant vectors for use with Agrobacterium and are constructed in the following manner. pTJS75kan is described in pTJS75, Schmidhauser & Helinski, J Bacteriol. 164: 446-455 (1985)] by digesting NarI to cleave the tetracycline resistance gene, followed by pUC4K with NPTII (Vieira & Messing, Gene 19: 259-268 (1982); Bevan et al., Nature 304: 184-187 (1983); McBride et al., Plant Molecular Biology 14: 266-276 (1990), to insert the AccI fragment. XhoI linkers were added to EcoRV fragments of pCIB7 containing left and right T-DNA boundaries, plant selectable nos / nptII chimeric genes, and pUC polylinkers (Rothstein et al., Gene 53: 153-161 (1987)). Ligation and cloning XhoI-digested fragments to SalI-digested pTJS75kan to prepare pCIB200 (see EP 0 332 104, Example 19). pCIB200 contains the following unique polylinker restriction sites: EcoRI, SstI, KpnI, BglII, XbaI and SalI. pCIB2001 is a derivative of pCIB200 prepared by inserting additional restriction sites into the polylinker. Intrinsic restriction sites in the polylinker of pCIB2001 are EcoRI, SstI, KpnI, BglII, XbaI, SalI, MluI, BcII, AvrII, ApaI, HpaI and StuI. In addition to these unique restriction sites, pCIB2001 is a plant and bacterial kanamycin selection gene, left and right T-DNA border genes for Agrobacterium-mediated transformation, E. coli. RK2-derived trfA agonist genes for migration between E. coli and other hosts, and also OriT and OriV agonist genes from RK2. The pCIB2001 polylinker is suitable for cloning plant expression cassettes containing their regulatory signals.

b. pCIB10 and its hygromycin selective derivatives

The double vector pCIB10 contains genes encoding kanamycin resistance for selection in plants and T-DNA right and left economic genes, including the broad host range plasmid pRK252. It can be replicated in both E. coli and Agrobacterium. Its construction is described in Rothstein et al. Gene 53: 153-161 (1987). Various derivatives of pCIB10 are constructed to include the genes of hygromycin B phosphotransferase described in Gritz et al., Gene 25: 179-188 (1983). These derivatives allow the selection of transgenic plant cells for hygromycin only (pCIB743) or for hygromycin and kanamycin (pCIB715, pCIB717).

2. Vectors Suitable for Non-Agrobacterium Transformation

Transformation without using Agrobacterium tumutation does not necessarily require a T-DNA sequence in the selected transformation vector, so that in addition to the vector described above containing the T-DNA sequence, a vector having the sequence deleted may be used. Transformation techniques that do not depend on Agrobacterium include transformation via particle bombardment, protoplast uptake (eg PEG and electropenetration) and microinjection. The choice of vector is highly dependent on the desired choice of the species to be transformed. The following describes common vectors suitable for non-Agrobacterium transformation.

a. pCIB3064

pCIB3064 is a vector derived from pUC that is suitable for direct gene transfer techniques with selection by the herbicide Basa (or phosphinothricin). Plasmid pCIB246 is E. coli. CaMV 35S promoter and CaMV 35S transcriptional terminator operably fused to the E. coli GUS gene and are described in PCT Publication No. WO 93/07278. The 35S promoter of the vector comprises two ATG sequences at 5 'of the starting site. These sites are mutated using standard PCR techniques in such a way that ATG is removed and restriction sites Ssp1 and PvuII are produced. The new restriction sites are 96 bp and 37 bp from the unique SalI site and 101 bp and 42 bp from the actual start site. The derivative of pCIB246 obtained is designated pCIB3025. The plasmid pCIB3060 is then prepared by digesting with SalI and SacI to remove the GUS gene from pCIB3025, blunting the ends and relinking. Plasmid pJIT82 was obtained from the John Innes Center, Norwich and cleaved a 400 bp SmaI fragment containing the bar gene from Streptomyces viridochromogenes and inserted into the HpaI site of pCIB3060. References: Thompson et al. EMBO J 6: 2519-2523 (1987). This gives pCIB3064 comprising a herbicide selection bar gene under CaMV 35S promoter and terminator control, an ampicillin resistance (for E. coli selection) gene and a polylinker with unique sites SphI, PstI, HindIII and BamHI. The vector is suitable for cloning a plant expression cassette containing its regulatory signal.

b. pSOG19 and pSOG35

pSOG35 is a selectable marker that confers resistance to methotrexate. Transformation vector using E. coli gene dihydrofolate reductase (DHFR). PCR is used to amplify the 35S promoter (approximately 800 bp), Intron 6 from the maize Adh1 gene (approximately 550 bp) and GUS unread leader sequence 18 bp from pSOG10. Also, this. The 250 bp fragment encoding the coli dihydrofolate reductase type II gene was amplified by PCR and these two PCR fragments were converted to SacI-PstI from pBI221 (Clontech), which includes the pUC19 vector backbone and nopaline synthase terminator. Integrate with the fragment. These fragments are aggregated to obtain pSOG19 containing a 35S promoter fused with an Intron 6 sequence, a GUS leader, a DHFR gene and a nopaline synthase terminator. The vector pSOG35 is prepared by replacing the GUS leader in pSOG19 with the leader sequence from maize lobular virus (MCMV). pSOG19 and pSOG35 contain the pUC gene for ampicillin resistance and have HindIII, SphI, PstI and EcoRI sites useful for cloning foreign substances.                 

3. Vectors Suitable for Chloroplast Transformation

To express the nucleotide sequence of the present invention in plant plastids, the plastiform transformation vector pPH143 (Example 36 of WO 97/32011) is used. The nucleotide sequence is inserted into pPH143 to replace the PROTOX coding sequence. This vector is then used for chromatin transformation and selection of transformants for spectinomycin resistance. Alternatively, the nucleotide sequence is inserted into pPH143 to replace the aadA gene. In this case the transformants are chosen to be resistant to PROTOX inhibitors.

Example 7: Transformation

Once the nucleic acid sequence of the invention is cloned into an expression system, it is transformed into plant cells. Methods for transforming and regenerating plants are well known in the art. For example, Ti plasmid vectors were used for delivery of foreign DNA and direct DNA uptake, liposomes, electropenetration, microinjection and microinjection. Bacteria from the genus Agrobacterium can also be used to transform plant cells. In the following, representative techniques for transforming both dicotyledonous and monocotyledonous plants and representative chromatin transformation techniques are described.

1. Transformation of Dicotyledonous Plants

Transformation techniques for dicotyledonous plants are well known in the art and include Agrobacterium-based techniques and techniques that do not require Agrobacterium. Non-Agrobacterium technology involves the incorporation of exogenous genetic material directly by the pigment or cell. This can be accomplished by PEG or electropenetration mediated absorption, particle impact-mediated delivery or microinjection. Examples of such techniques are described in Paszkowski et al., EMBO J 3: 271-2722 (1984), Potrykus et al., Mol. Gen. Genet. 199: 169-177 (1985), Reich et al., Biotechnology 4: 1001-1004 (1986), and Klein et al., Nature 327: 70-73 (1987). In each case the transformed cells are regenerated into complete plants using standard techniques known in the art.

Agrobacterium-mediated transformation is the preferred technique for the transformation of dicotyledonous plants because of its high transformation efficiency and widespread use in many different species. Agrobacterium transformation typically involves the transfer of a binary vector with foreign DNA of interest (eg pCIB 200 or pCIB 2001) to the appropriate Agrobacterium strain, wherein the transfer is a Ti co-resident of the host Agrobacterium strain. Complement of vir genes carried on plasmids or in chromosomes (eg strain CIB542 for pCIB200 and pCIB2001). Uknes et al. Plant Cell 5: 159-169 (1993). The delivery of the recombinant binary vector to Agrobacterium is a bacterium that has a recombinant binary vector. E. coli, an adjuvant E. coli with a plasmid such as pRK2013 and capable of transferring a recombinant binary vector to a target Agrobacterium strain. This is achieved using a triple parental mating procedure using E. coli strains. Alternatively, the recombinant binary vector can be delivered to Agrobacterium by DNA transformation. See Hofgen & Wilmitzer, Nucl. Acids Res. 16: 9877 (1988).

Transformation of target plant species by recombinant Agrobacterium is commonly associated with the coculture of Agrobacterium with explants from plants and according to protocols well known in the art. Transgenic tissues with antibiotic or herbicide resistance markers present between the binary plasmid T-DNA boundaries are regenerated on selectable media.

Another approach to transforming plant cells with genes involves pushing inactive or biologically active particles into plant tissues and cells. The technique is described in US Pat. Nos. 4,945,050, 5,036,006 and 5,100,792. In general, this process involves the penetration of inert or biologically active particles into the cell's outer surface and propagation into the cell under conditions effective for incorporation into the cell's interior. When inert particles are used, the vector can be introduced into cells by coating the particles with a vector containing the desired gene. Alternatively, the target cells may be surrounded by a vector such that the vector is delivered intracellularly through the trajectory of the particle. Biologically active particles, such as dried yeast cells, dried bacteria or bacteriophages, each containing DNA to be introduced, may be propagated into plant cell tissue.

2. Transformation of monocots

At present, transformation of most monocot species is also common. Preferred techniques involve direct delivery of genes into protoplasts and impacting particles into callus using PEG or electropenetration techniques. Transformation can be performed on a single DNA species or on multiple DNA species (ie cotransformation), both of which are suitable for use in the present invention. Cotransformation may have the advantage of avoiding complete vector construction and producing transgenic plants that are capable of removing selectable markers in the next generation because the selectable markers are not linked to the position of the gene of interest. However, a disadvantage with cotransformation is that individual DNA species are inserted into the genome at less than 100% frequency [Schocher et al. Biotechnology 4: 1093-1096 (1986)].

Patent applications EP 0 292 435, EP 0 392 225 and WO 93/07278 describe techniques for preparing callus and protoplasts from superior suburbs of maize, transformation and transformation of protoplasts using PEG or electropenetration. Regeneration of corn plants from converted protoplasts is described. Gordon-Kamm et al. (Plant Cell 2: 603-618 (1990)) and Fromm et al. (Biotechnology 8: 833-839 (1990)) describe corn from A188 using particle bombardment. Transformation techniques of the week are disclosed. In addition, WO 93/07278 and Koziel et al., Biotechnology 11: 194-200 (1993) describe transformation techniques of superior suburbs of corn using particle bombardment. The technique uses 1.5-2.5 mm long immature corn embryos and impacted PDS-1000He bioistic devices cut from corn eggs 14-15 days after pollination.

Transformation of rice can be performed by direct gene transfer techniques using protoplasts or by particle bombardment. Protoplast-mediated transformations have been described for Japonica varieties and Indica varieties. See Zhang et al. Plant Cell Rep 7: 379-384 (1988); Shimamoto et al. Nature 338: 274-277 (1989); Datta et al. Biotechnology 8: 736-740 (1990). In addition, both varieties can typically be transformed using particle bombardment. See Christou et al. Biotechnology 9: 957-962 (1991). WO 93/21335 also describes the transformation of rice through electropenetration.

Patent application EP 0 332 581 describes techniques for the generation, transformation and regeneration of rice and protoplasts. This technique allows the transformation of ducks and wheat. In addition, wheat transformation into cells of type C organ regenerative callus using particle bombardment (Vasil et al., Biotechnology 10: 667-674 (1992)) and of callus derived from immature embryos and immature embryos. Wheat transformation using particle bombardment [Vasil et al., Biotechnology 11: 1553-1558 (1993) and Weeks et al., Plant Physiol. 102: 1077-1084 (1993). However, a preferred wheat transformation technique involves the transformation of wheat by particle bombardment of immature embryos, and involves high sucrose or high maltose steps prior to gene transfer. Prior to the impact, to introduce somatic embryos, any number of embryos (0.75-1 mm in length) were subjected to 3% sucrose (Murashiga & Skoog, Physiologia Plantarum 15: 473-497 (1962)) and 3 mg / l 2 Smear in MS medium containing, 4-D and proceed in the dark. On the day of the impact, embryos are removed from the introduction medium and placed on an osmoticum (ie, introduction medium in which sucrose or maltose is added at the desired concentration, typically 15%). The embryos are separated for protoplasts for 2 to 3 hours and then shocked. Although not critical, 20 embryos per target plate are common. Appropriate gene containing plasmids (eg pCIB3064 or pSG35) are precipitated onto micrometer size gold particles using standard procedures. Each plate of embryos is fired using a DuPont Biolositics ^R helium device using a burst pressure of approximately 1000 psi and a standard 80 mesh screen. After the impact, the embryo is placed back in the dark to recover for about 24 hours (on the osmotic agent). After 24 hours, the embryos are removed from the osmotic agent and placed back on the infusion medium and left for about one month before regeneration. After about a month, embryonic implants with developing embryonic callus were regenerated medium (MS + 1 mg / s) containing additional appropriate selection agents (10 mg / l Baster for pCIB3064 and 2 mg / l methotrexate for pSOG35). 1 NAA, 5 mg / L GA). After about one month, the developing shoots are transferred to a large sterile container known as "GA7" containing 1/2 concentration of MS, 2% sucrose and the same concentration of the selection agent.

Transformation of monocots with Agrobacterium has also been described (WO 94/00977 and US Pat. No. 5,591,616; Both are incorporated herein by reference].

3. Transformation of Chromosome

Seeds of haploid tobacco (Nicotiana tabacum cv. 'Xanthi nc') are germinated 7 times per plate in a 1-inch circular arrangement on T agar medium, see Svab, Z. and Maliga, P. (1993) PNAS 90, 913-917, seeded together with 1 μm tungsten particles (M10, Biorad, Hercules, CA) coated with DNA from plasmids pPH143 and pPH145. 12 to 14 days after the shock is applied. After culturing for 2 days in shock medium, the leaves were cut and RMOP medium containing 500 μg / ml spectromycin dihydrochloride (Sigma, St. Louis, Mo., Svab). , Z., Hajdukiewicz, P. and Maliga, P. (1990). Three to eight weeks after the impact, the resistant shoots appearing under the bleached leaves are acloned in the same selectable medium to form callus, and the secondary shoots are isolated and acloned. Southern blotting for complete isolation of transformed chromatin genome copies (monotropic) in independent aclones (Sambrook et al., (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor) Evaluate using standard techniques. BamHI / EcoRI-digested whole cell DNA [Maettler, IJ (1987) Plant Mol Biol Reporter 5, 346-349] was isolated on 1% tris-borate (TBE) agarose gel and nylon membrane (Amersham) ) And then probe with a ³² P-labeled randomly primed DNA sequence corresponding to 0.7 kb BamHI / HindIII DNA fragment from pC8 containing a portion of the rps7 / 12 chromosomal targeting sequence. WO 98/11235). Monomeric shoots were treated with spectinomycin-containing MS / IBA medium [McBride, Ke et al. (1994) PNAS 91, 7301-7305, aseptically, and then transferred to the greenhouse.

Example 8: Sarcoma

The plant obtained through transformation with the nucleic acid sequence of the present invention may be any one of a wide variety of plant species, including monocotyledonous and dicotyledonous, but the plant used in the method of the present invention is preferably agriculturally It is selected from the list of important target crops. Expression of other properties important to production and quality with the genes of the present invention can be incorporated primarily into plants through breeding. Breeding approaches and techniques are known in the art. Welsh J. R., Fundamentals of Plant Genetics and Breeding, John Wiley & Sons, NY (1981); Crop Breeding, Wood D. R. (Ed.) American Society of Agronomy Madisons, Wisconsin (1983); Mayo O., The Theory of Plant Breeding 2nd Edition, Clarendon Press, Oxford (1987); Singh, D. P., Breeding for Resistance to Diseases and Insect Pests, Springer-Verlag, NY (1986); Wricke and Weber, Quantitative Genetics and Selection Plant Breeding, Walter de Gruyter and Co., Berlin (1986)].

The genetic properties engineered with the transgenic seeds and plants described above are transmitted by sexual reproduction or trophic growth and thus can be maintained and propagated in progeny plants. Generally such maintenance and propagation uses known agricultural methods developed for special purposes such as tilling, sowing or harvesting. Specialized processes such as hydroponic or greenhouse technology can also be applied. Since cultivated plants are vulnerable to competition by weed plants as well as attacks caused by insects or infections, measures must be taken to control yields by controlling weeds, plant diseases, insects, nematodes and other adverse conditions. This includes the application of pesticides such as herbicides, fungicides, gonadotropins, nematicides, aging agents and insecticides, as well as means such as soil cultivation or removal of weeds and infected plants.

In addition, the beneficial genetic properties of the transgenic plants and seeds according to the invention are characterized by improved properties, such as improved resistance to pests, herbicides or stresses, improved nutritional value, increased yields, or slaughter or degranulation. It can be used in plant breeding aimed at developing plants having an improved structure which causes less loss due to them. The various breeding stages are characterized by well-defined human interventions such as the selection of the strains to be crossed, the direct pollination of the parent or the selection of suitable offspring plants. Depending on the desired properties, different breeding means are taken. Related arts are well known in the art and include, but are not limited to, hybridization, cross-breeding, backcross breeding, multi-week breeding, species mixing, heterogeneous hybridization, dimer techniques, and the like. Hybridization techniques also include the use of mechanical, chemical or biological means to sterilize plants to produce male or female infertile plants. Passivated water using pollen of different strains of male sterile plants ensures that the genome of a male sterile but magnetic fertility plant will yield properties of both maternal strains uniformly. Thus, the transgenic seeds and plants according to the present invention may be used for the breeding of improved plant strains which, for example, increase the effectiveness of conventional methods such as herbicides or pesticide treatments or render the method unnecessary due to its modified genetic properties. Can be used. Or, because of the optimized gene "equipment", a novel with improved stress content that can produce a harvested product of higher quality compared to a product that may be insoluble against comparable developmental adverse conditions. Crops can be obtained.                 

Example 9: Seed Production

In seed production, the germination properties and uniformity of the seeds are essential production properties, while the germination properties and uniformity of the seeds harvested and marketed by the farmer are not important. Because crops are stored separately from other crops and weed seeds, it is difficult to control seed infectious diseases and produce seeds with good germination, they are very broad and well-received by seed producers working in the field of growing, conditioning and selling pure seeds. Defined seed production means have been developed. Therefore, it is customary for farmers to purchase guaranteed seeds that meet certain quality standards instead of using seeds harvested from their crops. The propagation material intended to be used as a seed is customarily treated with a protective coating comprising herbicides, insecticides, fungicides, fungicides, nematicides, fungicides or mixtures thereof. Protective coatings customarily used include compounds such as captan, carboxycin, thiram (TMTD ^R ), metallaxyl (Apron ^R ) and pyrimifos-methyl (Actellic ^R ). If desired, these compounds are formulated with carriers, surfactants or application-promoting adjuvants commonly used in the formulation art to protect against damage caused by bacterial, fungal or animal pests. Protective coatings may be applied by immersing the propagation material in a liquid formulation or by coating with a combined wet or dry formulation. Other methods of application are possible, such as treatment with shoots or berries.

Example 10 Corn Plant Analysis

Corn plants transformed with plasmids pNOV1436, pNOV1441 and pNOV1313 via Agrobacterium-mediated transformation show 100% mortality for European lighted moths and chestnut moths. ELISA data is presented below:

Example 11: Rice Plant Analysis

Rice plants transformed with plasmid pNOV1305 via Agrobacterium-mediated transformation show 100% mortality for European lighted moths and chestnut moths. ELISA data is presented below:

Example 12 Cabbage Plant Analysis

Cabbage plants transformed with plasmid pZU578 (SEQ ID NO: 17) via Agrobacterium-mediated transformation are tested for Plutella xylostella. The transgenic plants and the control plants are infected by transferring 16 larvae from four cabbage-rooted moth cultures (with cabbage plants) to four leaves of four larvae each with a paint brush. Infected plants are transferred to a 1 × 1 × 1 cage during the test period. Control plants include untransformed cabbage plants (sensitive controls) and untransformed cabbage plants sprayed with commercially available Bt insecticide Dipel (resistant control). The score (after 2 weeks) was-= no damage (or only small pore = resistance); + = Large hole in plant (= sensitive); ++ = Many large holes, the plant is severely damaged (= sensitive). Dipel plants always record a-score and the sensitivity control records a ++ score. Insect damage grades and ELISA data of the transgenic and control plants are shown below.

The above described embodiments are exemplary. The subject matter of the present invention is apparent to those skilled in the art, including many variations of the present invention. All such obvious and foreseeable variations are encompassed by the present invention.

<110> Syngenta Participations AG <120> Novel insecticidal toxins derived from Bacillus thuringiensis insecticidal       crystal proteins <130> 5-2000-054004-2 <140> <141> <150> US 60 / 227,956 <151> 2000-08-25 <160> 17 <170> KopatentIn 1.7 <210> 1 <211> 3579 <212> DNA <213> Artificial Sequences <220> <223> Description of Artificial Sequence: H04 with Cry1C       tail <220> <221> CDS (222) (1) .. (3579) <223> H04 with Cry1C tail <300> <303> Appl. Environ. Microbiol. <304> 62 <305> 5 <306> 1537-1543 <307> 1996 <300> <310> 5,736,131 <400> 1 atg gat aac aat ccg aac atc aat gaa tgc att cct tat aat tgt tta 48 Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu   1 5 10 15 agt aac cct gaa gta gaa gta tta ggt gga gaa aga ata gaa act ggt 96 Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly              20 25 30 tac acc cca atc gat att tcc ttg tcg cta acg caa ttt ctt ttg agt 144 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser          35 40 45 gaa ttt gtt ccc ggt gct gga ttt gtg tta gga cta gtt gat ata ata 192 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile      50 55 60 tgg gga att ttt ggt ccc tct caa tgg gac gca ttt ctt gta caa att 240 Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile  65 70 75 80 gaa cag tta att aac caa aga ata gaa gaa ttc gct agg aac caa gcc 288 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala                  85 90 95 att tct aga tta gaa gga cta agc aat ctt tat caa att tac gca gaa 336 Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu             100 105 110 tct ttt aga gag tgg gaa gca gat cct act aat cca gca tta aga gaa 384 Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu         115 120 125 gag atg cgt att caa ttc aat gac atg aac agt gcc ctt aca acc gct 432 Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala     130 135 140 att cct ctt ttt gca gtt caa aat tat caa gtt cct ctt tta tca gta 480 Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155 160 tat gtt caa gct gca aat tta cat tta tca gtt ttg aga gat gtt tca 528 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser                 165 170 175 gtg ttt gga caa agg tgg gga ttt gat gcc gcg act atc aat agt cgt 576 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg             180 185 190 tat aat gat tta act agg ctt att ggc aac tat aca gat cat gct gta 624 Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val         195 200 205 cgc tgg tac aat acg gga tta gag cgt gta tgg gga ccg gat tct aga 672 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg     210 215 220 gat tgg ata aga tat aat caa ttt aga aga gaa tta aca cta act gta 720 Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 tta gat atc gtt tct cta ttt ccg aac tat gat agt aga acg tat cca 768 Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro                 245 250 255 att cga aca gtt tcc caa tta aca aga gaa att tat aca aac cca gta 816 Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val             260 265 270 tta gaa aat ttt gat ggt agt ttt cga ggc tcg gct cag ggc ata gaa 864 Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu         275 280 285 gga agt att agg agt cca cat ttg atg gat ata ctt aac agt ata acc 912 Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr     290 295 300 atc tat acg gat gct cat aga gga gaa tat tat tgg tca ggg cat caa 960 Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln 305 310 315 320 ata atg gct tct cct gta ggg ttt tcg ggg cca gaa ttc act ttt ccg 1008 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro                 325 330 335 cta tat gga act atg gga aat gca gct cca caa caa cgt att gtt gct 1056 Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala             340 345 350 caa cta ggt cag ggc gtg tat aga aca tta tcg tcc act tta tat aga 1104 Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg         355 360 365 aga cct ttt aat ata ggg ata aat aat caa caa cta tct gtt ctt gac 1152 Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp     370 375 380 ggg aca gaa ttt gct tat gga acc tcc tca aat ttg cca tcc gct gta 1200 Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400 tac aga aaa agc gga acg gta gat tcg ctg gat gaa ata ccg cca cag 1248 Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln                 405 410 415 aat aac aac gtg cca cct agg caa gga ttt agt cat cga tta agc cat 1296 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His             420 425 430 gtt tca atg ttt cgt tca ggc ttt agt aat agt agt gta agt ata ata 1344 Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile         435 440 445 aga gct cct atg ttc tct tgg ata cat cgt agt gca act ctt aca aat 1392 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Thr Leu Thr Asn     450 455 460 aca att gat cca gag aga att aat caa ata cct tta gtg aaa gga ttt 1440 Thr Ile Asp Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe 465 470 475 480 aga gtt tgg ggg ggc acc tct gtc att aca gga cca gga ttt aca gga 1488 Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly                 485 490 495 ggg gat atc ctt cga aga aat acc ttt ggt gat ttt gta tct cta caa 1536 Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln             500 505 510 gtc aat att aat tca cca att acc caa aga tac cgt tta aga ttt cgt 1584 Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg         515 520 525 tac gct tcc agt agg gat gca cga gtt ata gta tta aca gga gcg gca 1632 Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala     530 535 540 tcc aca gga gtg gga ggc caa gtt agt gta aat atg cct ctt cag aaa 1680 Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys 545 550 555 560 act atg gaa ata ggg gag aac tta aca tct aga aca ttt aga tat acc 1728 Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr                 565 570 575 gat ttt agt aat cct ttt tca ttt aga gct aat cca gat ata att ggg 1776 Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly             580 585 590 ata agt gaa caa cct cta ttt ggt gca ggt tct att agt agc ggt gaa 1824 Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu         595 600 605 ctt tat ata gat aaa att gaa att att cta gca gat gca aca ttt gaa 1872 Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu     610 615 620 gca gaa tct gat tta gaa aga gca caa aag gcg gtg aat gcc ctg ttt 1920 Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Ala Leu Phe 625 630 635 640 act tct tcc aat caa atc ggg tta aaa acc gat gtg acg gat tat cat 1968 Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His                 645 650 655 att gat caa gta tcc aat tta gtg gat tgt tta tca gat gaa ttt tgt 2016 Ile Asp Gln Val Ser Asn Leu Val Asp Cys Leu Ser Asp Glu Phe Cys             660 665 670 ctg gat gaa aag cga gaa ttg tcc gag aaa gtc aaa cat gcg aag cga 2064 Leu Asp Glu Lys Arg Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg         675 680 685 ctc agt gat gag cgg aat tta ctt caa gat cca aac ttc aga ggg atc 2112 Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe Arg Gly Ile     690 695 700 aat aga caa cca gac cgt ggc tgg aga gga agt aca gat att acc atc 2160 Asn Arg Gln Pro Asp Arg Gly Trp Arg Gly Ser Thr Asp Ile Thr Ile 705 710 715 720 caa gga gga gat gac gta ttc aaa gag aat tac gtc aca cta ccg ggt 2208 Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Pro Gly                 725 730 735 acc gtt gat gag tgc tat cca acg tat tta tat cag aaa ata gat gag 2256 Thr Val Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu             740 745 750 tcg aaa tta aaa gct tat acc cgt tat gaa tta aga ggg tat atc gaa 2304 Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Glu Leu Arg Gly Tyr Ile Glu         755 760 765 gat agt caa gac tta gaa atc tat ttg atc cgt tac aat gca aaa cac 2352 Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His     770 775 780 gaa ata gta aat gtg cca ggc acg ggt tcc tta tgg ccg ctt tca gcc 2400 Glu Ile Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Leu Ser Ala 785 790 795 800 caa agt cca atc gga aag tgt gga gaa ccg aat cga tgc gcg cca cac 2448 Gln Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn Arg Cys Ala Pro His                 805 810 815 ctt gaa tgg aat cct gat cta gat tgt tcc tgc aga gac ggg gaa aaa 2496 Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg Asp Gly Glu Lys             820 825 830 tgt gca cat cat tcc cat cat ttc acc ttg gat att gat gtt gga tgt 2544 Cys Ala His His Ser His His Phe Thr Leu Asp Ile Asp Val Gly Cys         835 840 845 aca gac tta aat gag gac tta ggt gta tgg gtg ata ttc aag att aag 2592 Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe Lys Ile Lys     850 855 860 acg caa gat ggc cat gca aga cta ggg aat cta gag ttt ctc gaa gag 2640 Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu 865 870 875 880 aaa cca tta tta ggg gaa gca cta gct cgt gtg aaa aga gcg gag aag 2688 Lys Pro Leu Leu Gly Glu Ala Leu Ala Arg Val Lys Arg Ala Glu Lys                 885 890 895 aag tgg aga gac aaa cga gag aaa ctg cag ttg gaa aca aat att gtt 2736 Lys Trp Arg Asp Lys Arg Glu Lys Leu Gln Leu Glu Thr Asn Ile Val             900 905 910 tat aaa gag gca aaa gaa tct gta gat gct tta ttt gta aac tct caa 2784 Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val Asn Ser Gln         915 920 925 tat gat aga tta caa gtg gat acg aac atc gcg atg att cat gcg gca 2832 Tyr Asp Arg Leu Gln Val Asp Thr Asn Ile Ala Met Ile His Ala Ala     930 935 940 gat aaa cgc gtt cat aga atc cgg gaa gcg tat ctg cca gag ttg tct 2880 Asp Lys Arg Val His Arg Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser 945 950 955 960 gtg att cca ggt gtc aat gcg gcc att ttc gaa gaa tta gag gga cgt 2928 Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu Glu Gly Arg                 965 970 975 att ttt aca gcg tat tcc tta tat gat gcg aga aat gtc att aaa aat 2976 Ile Phe Thr Ala Tyr Ser Leu Tyr Asp Ala Arg Asn Val Ile Lys Asn             980 985 990 ggc gat ttc aat aat ggc tta tta tgc tgg aac gtg aaa ggt cat gta 3024 Gly Asp Phe Asn Asn Gly Leu Leu Cys Trp Asn Val Lys Gly His Val         995 1000 1005 gat gta gaa gag caa aac aac cac cgt tcg gtc ctt gtt atc cca gaa 3072 Asp Val Glu Glu Gln Asn Asn His Arg Ser Val Leu Val Ile Pro Glu    1010 1015 1020 tgg gag gca gaa gtg tca caa gag gtt cgt gtc tgt cca ggt cgt ggc 3120 Trp Glu Ala Glu Val Ser Gln Glu Val Arg Val Cys Pro Gly Arg Gly 1025 1030 1035 1040 tat atc ctt cgt gtc aca gca tat aaa gag gga tat gga gag ggc tgc 3168 Tyr Ile Leu Arg Val Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys                1045 1050 1055 gta acg atc cat gag atc gaa gac aat aca gac gaa ctg aaa ttc agc 3216 Val Thr Ile His Glu Ile Glu Asp Asn Thr Asp Glu Leu Lys Phe Ser            1060 1065 1070 aac tgt gta gaa gag gaa gta tat cca aac aac aca gta acg tgt aat 3264 Asn Cys Val Glu Glu Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn        1075 1080 1085 aat tat act ggg act caa gaa gaa tat gag ggt acg tac act tct cgt 3312 Asn Tyr Thr Gly Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg    1090 1095 1100 aat caa gga tat gac gaa gcc tat ggt aat aac cct tcc gta cca gct 3360 Asn Gln Gly Tyr Asp Glu Ala Tyr Gly Asn Asn Pro Ser Val Pro Ala 1105 1110 1115 1120 gat tac gct tca gtc tat gaa gaa aaa tcg tat aca gat gga cga aga 3408 Asp Tyr Ala Ser Val Tyr Glu Glu Lys Ser Tyr Thr Asp Gly Arg Arg                1125 1130 1135 gag aat cct tgt gaa tct aac aga ggc tat ggg gat tac aca cca cta 3456 Glu Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu            1140 1145 1150 ccg gct ggt tat gta aca aag gat tta gag tac ttc cca gag acc gat 3504 Pro Ala Gly Tyr Val Thr Lys Asp Leu Glu Tyr Phe Pro Glu Thr Asp        1155 1160 1165 aag gta tgg att gag atc gga gaa aca gaa gga aca ttc atc gtg gat 3552 Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val Asp    1170 1175 1180 agc gtg gaa tta ctc ctt atg gag gaa 3579 Ser Val Glu Leu Leu Leu Met Glu Glu 1185 1190 <210> 2 <211> 1193 <212> PRT <213> Artificial Sequence <223> Description of Artificial Sequence: H04 with Cry1C       tail <400> 2 Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu   1 5 10 15 Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly              20 25 30 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser          35 40 45 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile      50 55 60 Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile  65 70 75 80 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala                  85 90 95 Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu             100 105 110 Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu         115 120 125 Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala     130 135 140 Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155 160 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser                 165 170 175 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg             180 185 190 Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val         195 200 205 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg     210 215 220 Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro                 245 250 255 Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val             260 265 270 Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu         275 280 285 Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr     290 295 300 Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln 305 310 315 320 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro                 325 330 335 Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala             340 345 350 Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg         355 360 365 Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp     370 375 380 Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400 Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln                 405 410 415 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His             420 425 430 Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile         435 440 445 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Thr Leu Thr Asn     450 455 460 Thr Ile Asp Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe 465 470 475 480 Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly                 485 490 495 Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln             500 505 510 Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg         515 520 525 Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala     530 535 540 Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys 545 550 555 560 Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr                 565 570 575 Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly             580 585 590 Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu         595 600 605 Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu     610 615 620 Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Ala Leu Phe 625 630 635 640 Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His                 645 650 655 Ile Asp Gln Val Ser Asn Leu Val Asp Cys Leu Ser Asp Glu Phe Cys             660 665 670 Leu Asp Glu Lys Arg Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg         675 680 685 Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe Arg Gly Ile     690 695 700 Asn Arg Gln Pro Asp Arg Gly Trp Arg Gly Ser Thr Asp Ile Thr Ile 705 710 715 720 Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Pro Gly                 725 730 735 Thr Val Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Lys Ile Asp Glu             740 745 750 Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Glu Leu Arg Gly Tyr Ile Glu         755 760 765 Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His     770 775 780 Glu Ile Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Leu Ser Ala 785 790 795 800 Gln Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn Arg Cys Ala Pro His                 805 810 815 Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg Asp Gly Glu Lys             820 825 830 Cys Ala His His Ser His His Phe Thr Leu Asp Ile Asp Val Gly Cys         835 840 845 Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe Lys Ile Lys     850 855 860 Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu 865 870 875 880 Lys Pro Leu Leu Gly Glu Ala Leu Ala Arg Val Lys Arg Ala Glu Lys                 885 890 895 Lys Trp Arg Asp Lys Arg Glu Lys Leu Gln Leu Glu Thr Asn Ile Val             900 905 910 Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val Asn Ser Gln         915 920 925 Tyr Asp Arg Leu Gln Val Asp Thr Asn Ile Ala Met Ile His Ala Ala     930 935 940 Asp Lys Arg Val His Arg Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser 945 950 955 960 Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu Glu Gly Arg                 965 970 975 Ile Phe Thr Ala Tyr Ser Leu Tyr Asp Ala Arg Asn Val Ile Lys Asn             980 985 990 Gly Asp Phe Asn Asn Gly Leu Leu Cys Trp Asn Val Lys Gly His Val         995 1000 1005 Asp Val Glu Glu Gln Asn Asn His Arg Ser Val Leu Val Ile Pro Glu    1010 1015 1020 Trp Glu Ala Glu Val Ser Gln Glu Val Arg Val Cys Pro Gly Arg Gly 025 1030 1035 1040 Tyr Ile Leu Arg Val Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys                1045 1050 1055 Val Thr Ile His Glu Ile Glu Asp Asn Thr Asp Glu Leu Lys Phe Ser            1060 1065 1070 Asn Cys Val Glu Glu Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn        1075 1080 1085 Asn Tyr Thr Gly Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg    1090 1095 1100 Asn Gln Gly Tyr Asp Glu Ala Tyr Gly Asn Asn Pro Ser Val Pro Ala 105 1110 1115 1120 Asp Tyr Ala Ser Val Tyr Glu Glu Lys Ser Tyr Thr Asp Gly Arg Arg                1125 1130 1135 Glu Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu            1140 1145 1150 Pro Ala Gly Tyr Val Thr Lys Asp Leu Glu Tyr Phe Pro Glu Thr Asp        1155 1160 1165 Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val Asp    1170 1175 1180 Ser Val Glu Leu Leu Leu Met Glu Glu 185 1190 <210> 3 <211> 1896 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic gene       encoding the toxin portion of H04 without a tail <220> <221> CDS (222) (1) .. (1896) <223> H04 toxin portion without a tail <400> 3 atg gac aac aac ccc aac atc aac gag tgc atc ccc tac aac tgc ctg 48 Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu   1 5 10 15 agc aac ccc gag gtg gag gtg ctg ggc ggc gag cgc atc gag acc ggc 96 Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly              20 25 30 tac acc ccc atc gac atc agc ctg agc ctg acc cag ttc ctg ctg agc 144 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser          35 40 45 gag ttc gtg ccc ggc gcc ggc ttc gtg ctg ggc ctg gtg gac atc atc 192 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile      50 55 60 tgg ggc atc ttc ggc ccc agc cag tgg gac gcc ttc ctg gtg cag atc 240 Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile  65 70 75 80 gag cag ttg ata aac caa cgc ata gag gaa ttc gcc cgc aac cag gcc 288 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala                  85 90 95 atc agc cgc ctg gag ggc ctg agc aac ctg tac caa atc tac gcc gag 336 Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu             100 105 110 agc ttc cgc gag tgg gag gcc gac ccc acc aac ccc gcc ctg cgc gag 384 Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu         115 120 125 gag atg cgc atc cag ttc aac gac atg aac agc gcc ctg acc acc gcc 432 Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala     130 135 140 atc ccc ctg ttc gcc gtg cag aac tac cag gtg ccc ctg ctg agc gtg 480 Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155 160 tac gtg cag gcc gcc aac ctg cac ctg agc gtg ctg cgc gac gtc agc 528 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser                 165 170 175 gtg ttc ggc cag cgc tgg ggc ttc gac gcc gcc acc atc aac agc cgc 576 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg             180 185 190 tac aac gac ctg acc cgc ctg atc ggc aac tac acc gac cac gcc gtg 624 Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val         195 200 205 cgc tgg tac aac acc ggc ctg gag cgc gtg tgg ggt ccc gac agc cgc 672 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg     210 215 220 gac tgg atc agg tac aac cag ttc cgc cgc gag ctg acc ctg acc gtg 720 Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 ctg gac atc gtg agc ctg ttc ccc aac tac gac agc cgc acc tac ccc 768 Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro                 245 250 255 atc cgc acc gtg agc cag ctg acc cgc gag att tac acc aac ccc gtg 816 Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val             260 265 270 ctg gag aac ttc gac ggc agc ttc cgc ggc agc gcc cag ggc atc gag 864 Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu         275 280 285 ggc agc atc cgc agc ccc cac ctg atg gac atc ctg aac agc atc acc 912 Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr     290 295 300 atc tac acc gac gcc cac cgc ggc gag tac tac tgg agc ggc cac cag 960 Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln 305 310 315 320 atc atg gcc agc ccc gtc ggc ttc agc ggc ccc gag ttc acc ttc ccc 1008 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro                 325 330 335 ctg tac ggc acc atg ggc aac gct gca cct cag cag cgc atc gtg gca 1056 Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala             340 345 350 cag ctg ggc cag gga gtg tac cgc acc ctg agc agc acc ctg tac cgt 1104 Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg         355 360 365 cga cct ttc aac atc ggc atc aac aac cag cag ctg agc gtg ctg gac 1152 Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp     370 375 380 ggc acc gag ttc gcc tac ggc acc agc agc aac ctg ccc agc gcc gtg 1200 Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400 tac cgc aag agc ggc acc gtg gac agc ctg gac gag atc ccc cct cag 1248 Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln                 405 410 415 aac aac aac gtg cca cct cga cag ggc ttc agc cac cgt ctg agc cac 1296 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His             420 425 430 gtg agc atg ttc cgc agt ggc ttc agc aac agc agc gtg agc atc atc 1344 Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile         435 440 445 cgt gca ccc atg ttc agc tgg att cac cgc agc gcc acc ctg acc aac 1392 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Thr Leu Thr Asn     450 455 460 acc atc gac ccc gag cgc atc aac cag atc ccc ctg gtg aag ggc ttc 1440 Thr Ile Asp Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe 465 470 475 480 cgg gtg tgg ggc ggc acc agc gtg atc acc ggc ccc ggc ttc acc gga 1488 Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly                 485 490 495 ggc gac atc ctg cgc aga aac acc ttc ggc gac ttc gtg agc ctg cag 1536 Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln             500 505 510 gtg aac atc aac agc ccc atc acc cag cgt tac cgc ctg cgc ttc cgc 1584 Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg         515 520 525 tac gcc agc agc cgc gac gcc cgt gtg atc gtg ctg act ggc gcc gct 1632 Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala     530 535 540 agc acc ggt gtg ggc ggt cag gtg agc gtg aac atg ccc ctg cag aag 1680 Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys 545 550 555 560 act atg gag atc ggc gag aac ctg act agt cgc acc ttc cgc tac acc 1728 Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr                 565 570 575 gac ttc agc aac ccc ttc agc ttc cgc gcc aac ccc gac atc atc ggc 1776 Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly             580 585 590 atc agc gag cag ccc ctg ttc ggt gcc ggc agc atc agc agc ggc gag 1824 Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu         595 600 605 ctg tac atc gac aag atc gag atc atc ctg gcc gac gcc acc ttc gag 1872 Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu     610 615 620 gcc gag agc gac ctg gag cgc taa 1896 Ala Glu Ser Asp Leu Glu Arg 625 630 <210> 4 <211> 631 <212> PRT <213> Artificial Sequence <223> Description of Artificial Sequence: synthetic gene       encoding the toxin portion of H04 without a tail <400> 4 Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu   1 5 10 15 Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly              20 25 30 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser          35 40 45 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile      50 55 60 Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile  65 70 75 80 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala                  85 90 95 Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu             100 105 110 Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu         115 120 125 Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala     130 135 140 Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155 160 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser                 165 170 175 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg             180 185 190 Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val         195 200 205 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg     210 215 220 Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro                 245 250 255 Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val             260 265 270 Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu         275 280 285 Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr     290 295 300 Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln 305 310 315 320 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro                 325 330 335 Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala             340 345 350 Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg         355 360 365 Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp     370 375 380 Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400 Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln                 405 410 415 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His             420 425 430 Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile         435 440 445 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Thr Leu Thr Asn     450 455 460 Thr Ile Asp Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe 465 470 475 480 Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly                 485 490 495 Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln             500 505 510 Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg         515 520 525 Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala     530 535 540 Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys 545 550 555 560 Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr                 565 570 575 Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly             580 585 590 Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu         595 600 605 Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu     610 615 620 Ala Glu Ser Asp Leu Glu Arg 625 630 <210> 5 <211> 3582 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic gene       encoding H04 with full-length Cry1Ab tail <220> <221> CDS (222) (1) .. (3582) <223> H04 with full-length Cry1Ab tail <400> 5 atg gac aac aac ccc aac atc aac gag tgc atc ccc tac aac tgc ctg 48 Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu   1 5 10 15 agc aac ccc gag gtg gag gtg ctg ggc ggc gag cgc atc gag acc ggc 96 Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly              20 25 30 tac acc ccc atc gac atc agc ctg agc ctg acc cag ttc ctg ctg agc 144 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser          35 40 45 gag ttc gtg ccc ggc gcc ggc ttc gtg ctg ggc ctg gtg gac atc atc 192 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile      50 55 60 tgg ggc atc ttc ggc ccc agc cag tgg gac gcc ttc ctg gtg cag atc 240 Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile  65 70 75 80 gag cag ttg ata aac caa cgc ata gag gaa ttc gcc cgc aac cag gcc 288 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala                  85 90 95 atc agc cgc ctg gag ggc ctg agc aac ctg tac caa atc tac gcc gag 336 Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu             100 105 110 agc ttc cgc gag tgg gag gcc gac ccc acc aac ccc gcc ctg cgc gag 384 Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu         115 120 125 gag atg cgc atc cag ttc aac gac atg aac agc gcc ctg acc acc gcc 432 Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala     130 135 140 atc ccc ctg ttc gcc gtg cag aac tac cag gtg ccc ctg ctg agc gtg 480 Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155 160 tac gtg cag gcc gcc aac ctg cac ctg agc gtg ctg cgc gac gtc agc 528 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser                 165 170 175 gtg ttc ggc cag cgc tgg ggc ttc gac gcc gcc acc atc aac agc cgc 576 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg             180 185 190 tac aac gac ctg acc cgc ctg atc ggc aac tac acc gac cac gcc gtg 624 Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val         195 200 205 cgc tgg tac aac acc ggc ctg gag cgc gtg tgg ggt ccc gac agc cgc 672 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg     210 215 220 gac tgg atc agg tac aac cag ttc cgc cgc gag ctg acc ctg acc gtg 720 Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 ctg gac atc gtg agc ctg ttc ccc aac tac gac agc cgc acc tac ccc 768 Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro                 245 250 255 atc cgc acc gtg agc cag ctg acc cgc gag att tac acc aac ccc gtg 816 Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val             260 265 270 ctg gag aac ttc gac ggc agc ttc cgc ggc agc gcc cag ggc atc gag 864 Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu         275 280 285 ggc agc atc cgc agc ccc cac ctg atg gac atc ctg aac agc atc acc 912 Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr     290 295 300 atc tac acc gac gcc cac cgc ggc gag tac tac tgg agc ggc cac cag 960 Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln 305 310 315 320 atc atg gcc agc ccc gtc ggc ttc agc ggc ccc gag ttc acc ttc ccc 1008 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro                 325 330 335 ctg tac ggc acc atg ggc aac gct gca cct cag cag cgc atc gtg gca 1056 Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala             340 345 350 cag ctg ggc cag gga gtg tac cgc acc ctg agc agc acc ctg tac cgt 1104 Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg         355 360 365 cga cct ttc aac atc ggc atc aac aac cag cag ctg agc gtg ctg gac 1152 Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp     370 375 380 ggc acc gag ttc gcc tac ggc acc agc agc aac ctg ccc agc gcc gtg 1200 Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400 tac cgc aag agc ggc acc gtg gac agc ctg gac gag atc ccc cct cag 1248 Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln                 405 410 415 aac aac aac gtg cca cct cga cag ggc ttc agc cac cgt ctg agc cac 1296 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His             420 425 430 gtg agc atg ttc cgc agt ggc ttc agc aac agc agc gtg agc atc atc 1344 Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile         435 440 445 cgt gca ccc atg ttc agc tgg att cac cgc agc gcc acc ctg acc aac 1392 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Thr Leu Thr Asn     450 455 460 acc atc gac ccc gag cgc atc aac cag atc ccc ctg gtg aag ggc ttc 1440 Thr Ile Asp Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe 465 470 475 480 cgg gtg tgg ggc ggc acc agc gtg atc acc ggc ccc ggc ttc acc gga 1488 Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly                 485 490 495 ggc gac atc ctg cgc aga aac acc ttc ggc gac ttc gtg agc ctg cag 1536 Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln             500 505 510 gtg aac atc aac agc ccc atc acc cag cgt tac cgc ctg cgc ttc cgc 1584 Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg         515 520 525 tac gcc agc agc cgc gac gcc cgt gtg atc gtg ctg act ggc gcc gct 1632 Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala     530 535 540 agc acc ggt gtg ggc ggt cag gtg agc gtg aac atg ccc ctg cag aag 1680 Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys 545 550 555 560 act atg gag atc ggc gag aac ctg act agt cgc acc ttc cgc tac acc 1728 Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr                 565 570 575 gac ttc agc aac ccc ttc agc ttc cgc gcc aac ccc gac atc atc ggc 1776 Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly             580 585 590 atc agc gag cag ccc ctg ttc ggt gcc ggc agc atc agc agc ggc gag 1824 Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu         595 600 605 ctg tac atc gac aag atc gag atc atc ctg gcc gac gcc acc ttc gag 1872 Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu     610 615 620 gcc gag agc gac ctg gag cgc gcc cag aag gcc gtg aac gcc ctg ttc 1920 Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Ala Leu Phe 625 630 635 640 acc agc agc aac cag atc ggc ctg aag acc gac gtg acc gac tac cac 1968 Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His                 645 650 655 atc gac cag gtg agc aac ctg gtg gac tgc tta agc gac gag ttc tgc 2016 Ile Asp Gln Val Ser Asn Leu Val Asp Cys Leu Ser Asp Glu Phe Cys             660 665 670 ctg gac gag aag aag gag ctg agc gag aag gtg aag cac gcc aag cgc 2064 Leu Asp Glu Lys Lys Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg         675 680 685 ctg agc gac gag cgc aac ctg ctg cag gac ccc aac ttc cgc ggc atc 2112 Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe Arg Gly Ile     690 695 700 aac cgc cag ctg gac cgc ggc tgg cga ggc agc acc gat atc acc atc 2160 Asn Arg Gln Leu Asp Arg Gly Trp Arg Gly Ser Thr Asp Ile Thr Ile 705 710 715 720 cag ggc ggc gac gac gtg ttc aag gag aac tac gtg acc ctg cag ggc 2208 Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Gln Gly                 725 730 735 acc ttc gac gag tgc tac ccc acc tac ctg tac cag ccg atc gac gag 2256 Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Pro Ile Asp Glu             740 745 750 agc aag ctg aag gcc tac acc cgc tac cag ctg cgc ggc tac atc gag 2304 Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg Gly Tyr Ile Glu         755 760 765 gac agc cag gac ctg gaa atc tac ctg atc cgc tac aac gcg aag cac 2352 Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His     770 775 780 gag acc gtg aac gtg ccc ggc acc ggc agc ctg tgg ccc ccg agc gcc 2400 Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Pro Ser Ala 785 790 795 800 ccc agc ccc atc ggc aag tgc ggg gag ccg aat cga tgc gct ccg cac 2448 Pro Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn Arg Cys Ala Pro His                 805 810 815 ctg gag tgg aac ccg gac cta gac tgc agc tgc agg gac ggg gag aag 2496 Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg Asp Gly Glu Lys             820 825 830 tgc gcc cac cac agc cac cac ttc agc ctg gac atc gac gtg ggc tgc 2544 Cys Ala His His Ser His His Phe Ser Leu Asp Ile Asp Val Gly Cys         835 840 845 acc gac ctg aac gag gac ctg ggc gtg tgg gtg atc ttc aag atc aag 2592 Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe Lys Ile Lys     850 855 860 acc cag gac ggc cac gcc cgc ctg ggc aat cta gag ttc ctg gag gag 2640 Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu 865 870 875 880 aag ccc ctg gtg ggc gag gcc ctg gcc cgc gtg aag cgt gct gag aag 2688 Lys Pro Leu Val Gly Glu Ala Leu Ala Arg Val Lys Arg Ala Glu Lys                 885 890 895 aag tgg cgc gac aag cgc gag aag ctg gag tgg gag acc aac atc gtg 2736 Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu Trp Glu Thr Asn Ile Val             900 905 910 tac aag gag gcc aag gag agc gtg gac gcc ctg ttc gtg aac agc cag 2784 Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val Asn Ser Gln         915 920 925 tac gac cgc ctg cag gcc gac acc aac atc gcc atg atc cac gcc gcc 2832 Tyr Asp Arg Leu Gln Ala Asp Thr Asn Ile Ala Met Ile His Ala Ala     930 935 940 gac aag cgc gtg cac agc att cgc gag gcc tac ctg ccc gag ctg agc 2880 Asp Lys Arg Val His Ser Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser 945 950 955 960 gtg atc ccc ggt gtg aac gcc gcc atc ttc gag gaa ctc gag ggc cgc 2928 Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu Glu Gly Arg                 965 970 975 atc ttc acc gcc ttc agc ctg tac gac gcc cgc aac gtg atc aag aac 2976 Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala Arg Asn Val Ile Lys Asn             980 985 990 ggc gac ttc aac aac ggc ctg agc tgc tgg aac gtg aag ggc cac gtg 3024 Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp Asn Val Lys Gly His Val         995 1000 1005 gac gtg gag gag cag aac aac cac cgc agc gtg ctg gtg gtg ccc gag 3072 Asp Val Glu Glu Gln Asn Asn His Arg Ser Val Leu Val Val Pro Glu    1010 1015 1020 tgg gag gcc gag gtg agc cag gag gtg cgc gtg tgc ccc ggc cgc ggc 3120 Trp Glu Ala Glu Val Ser Gln Glu Val Arg Val Cys Pro Gly Arg Gly 1025 1030 1035 1040 tac atc ctg cgc gtg acc gcc tac aag gag ggc tac ggc gag ggc tgc 3168 Tyr Ile Leu Arg Val Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys                1045 1050 1055 gtg acc atc cac gag atc gag aac aac acc gac gag ctc aag ttc agc 3216 Val Thr Ile His Glu Ile Glu Asn Asn Thr Asp Glu Leu Lys Phe Ser            1060 1065 1070 aac tgc gtg gag gag gag gtt tac ccc aac aac acc gtg acc tgc aac 3264 Asn Cys Val Glu Glu Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn        1075 1080 1085 gac tac acc gcg acc cag gag gag tac gaa ggc acc tac acc tct cgc 3312 Asp Tyr Thr Ala Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg    1090 1095 1100 aac agg ggt tac gac ggc gcc tac gag tcc aac agc tcc gtg cca gct 3360 Asn Arg Gly Tyr Asp Gly Ala Tyr Glu Ser Asn Ser Ser Val Pro Ala 1105 1110 1115 1120 gac tac gcc agc gcc cac gag gag aaa gcc tac acc gac ggt aga cgc 3408 Asp Tyr Ala Ser Ala His Glu Glu Lys Ala Tyr Thr Asp Gly Arg Arg                1125 1130 1135 gac aac cca tgt gag agc aac aga ggc tac ggc gac tac acc ccc ctg 3456 Asp Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu            1140 1145 1150 ccc gct gga tac gtg acc aag gag ctg gag tac ttc ccc gag acc gac 3504 Pro Ala Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr Asp        1155 1160 1165 aag gtg tgg atc gag att ggc gag acc gag ggc acc ttc atc gtg gac 3552 Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val Asp    1170 1175 1180 agc gtg gag ctg ctg ctg atg gag gag tag 3582 Ser Val Glu Leu Leu Leu Met Glu Glu 1185 1190 <210> 6 <211> 1193 <212> PRT <213> Artificial Sequence <223> Description of Artificial Sequence: synthetic gene       encoding H04 with full-length Cry1Ab tail <400> 6 Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu   1 5 10 15 Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly              20 25 30 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser          35 40 45 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile      50 55 60 Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile  65 70 75 80 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala                  85 90 95 Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu             100 105 110 Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu         115 120 125 Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala     130 135 140 Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155 160 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser                 165 170 175 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg             180 185 190 Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val         195 200 205 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg     210 215 220 Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro                 245 250 255 Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val             260 265 270 Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu         275 280 285 Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr     290 295 300 Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln 305 310 315 320 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro                 325 330 335 Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala             340 345 350 Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg         355 360 365 Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp     370 375 380 Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400 Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln                 405 410 415 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His             420 425 430 Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile         435 440 445 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Thr Leu Thr Asn     450 455 460 Thr Ile Asp Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe 465 470 475 480 Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly                 485 490 495 Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln             500 505 510 Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg         515 520 525 Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala     530 535 540 Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys 545 550 555 560 Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr                 565 570 575 Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly             580 585 590 Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu         595 600 605 Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu     610 615 620 Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Ala Leu Phe 625 630 635 640 Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His                 645 650 655 Ile Asp Gln Val Ser Asn Leu Val Asp Cys Leu Ser Asp Glu Phe Cys             660 665 670 Leu Asp Glu Lys Lys Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg         675 680 685 Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe Arg Gly Ile     690 695 700 Asn Arg Gln Leu Asp Arg Gly Trp Arg Gly Ser Thr Asp Ile Thr Ile 705 710 715 720 Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Gln Gly                 725 730 735 Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Pro Ile Asp Glu             740 745 750 Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg Gly Tyr Ile Glu         755 760 765 Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His     770 775 780 Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Pro Ser Ala 785 790 795 800 Pro Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn Arg Cys Ala Pro His                 805 810 815 Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg Asp Gly Glu Lys             820 825 830 Cys Ala His His Ser His His Phe Ser Leu Asp Ile Asp Val Gly Cys         835 840 845 Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe Lys Ile Lys     850 855 860 Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu 865 870 875 880 Lys Pro Leu Val Gly Glu Ala Leu Ala Arg Val Lys Arg Ala Glu Lys                 885 890 895 Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu Trp Glu Thr Asn Ile Val             900 905 910 Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val Asn Ser Gln         915 920 925 Tyr Asp Arg Leu Gln Ala Asp Thr Asn Ile Ala Met Ile His Ala Ala     930 935 940 Asp Lys Arg Val His Ser Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser 945 950 955 960 Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu Glu Gly Arg                 965 970 975 Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala Arg Asn Val Ile Lys Asn             980 985 990 Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp Asn Val Lys Gly His Val         995 1000 1005 Asp Val Glu Glu Gln Asn Asn His Arg Ser Val Leu Val Val Pro Glu    1010 1015 1020 Trp Glu Ala Glu Val Ser Gln Glu Val Arg Val Cys Pro Gly Arg Gly 1025 1030 1035 1040 Tyr Ile Leu Arg Val Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys                1045 1050 1055 Val Thr Ile His Glu Ile Glu Asn Asn Thr Asp Glu Leu Lys Phe Ser            1060 1065 1070 Asn Cys Val Glu Glu Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn        1075 1080 1085 Asp Tyr Thr Ala Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg    1090 1095 1100 Asn Arg Gly Tyr Asp Gly Ala Tyr Glu Ser Asn Ser Ser Val Pro Ala 1105 1110 1115 1120 Asp Tyr Ala Ser Ala His Glu Glu Lys Ala Tyr Thr Asp Gly Arg Arg                1125 1130 1135 Asp Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu            1140 1145 1150 Pro Ala Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr Asp        1155 1160 1165 Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val Asp    1170 1175 1180 Ser Val Glu Leu Leu Leu Met Glu Glu 1185 1190 <210> 7 <211> 3582 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic gene       encoding H04 with full-length Cry1Ab tail <220> <221> CDS (222) (1) .. (3582) <223> H04 with full-length Cry1Ab tail <400> 7 atg gac aac aac ccc aac atc aac gag tgc atc ccc tac aac tgc ctg 48 Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu   1 5 10 15 agc aac ccc gag gtg gag gtg ctg ggc ggc gag cgc atc gag acc ggc 96 Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly              20 25 30 tac acc ccc atc gac atc agc ctg agc ctg acc cag ttc ctg ctg agc 144 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser          35 40 45 gag ttc gtg ccc ggc gcc ggc ttc gtg ctg ggc ctg gtg gac atc atc 192 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile      50 55 60 tgg ggc atc ttc ggc ccc agc cag tgg gac gcc ttc ctg gtg cag atc 240 Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile  65 70 75 80 gag cag ttg ata aac caa cgc ata gag gaa ttc gcc cgc aac cag gcc 288 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala                  85 90 95 atc agc cgc ctg gag ggc ctg agc aac ctg tac caa atc tac gcc gag 336 Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu             100 105 110 agc ttc cgc gag tgg gag gcc gac ccc acc aac ccc gcc ctg cgc gag 384 Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu         115 120 125 gag atg cgc atc cag ttc aac gac atg aac agc gcc ctg acc acc gcc 432 Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala     130 135 140 atc ccc ctg ttc gcc gtg cag aac tac cag gtg ccc ctg ctg agc gtg 480 Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155 160 tac gtg cag gcc gcc aac ctg cac ctg agc gtg ctg cgc gac gtc agc 528 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser                 165 170 175 gtg ttc ggc cag cgc tgg ggc ttc gac gcc gcc acc atc aac agc cgc 576 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg             180 185 190 tac aac gac ctg acc cgc ctg atc ggc aac tac acc gac cac gcc gtg 624 Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val         195 200 205 cgc tgg tac aac acc ggc ctg gag cgc gtg tgg ggt ccc gac agc cgc 672 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg     210 215 220 gac tgg atc agg tac aac cag ttc cgc cgc gag ctg acc ctg acc gtg 720 Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 ctg gac atc gtg agc ctg ttc ccc aac tac gac agc cgc acc tac ccc 768 Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro                 245 250 255 atc cgc acc gtg agc cag ctg acc cgc gag att tac acc aac ccc gtg 816 Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val             260 265 270 ctg gag aac ttc gac ggc agc ttc cgc ggc agc gcc cag ggc atc gag 864 Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu         275 280 285 ggc agc atc cgc agc ccc cac ctg atg gac atc ctg aac agc atc acc 912 Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr     290 295 300 atc tac acc gac gcc cac cgc ggc gag tac tac tgg agc ggc cac cag 960 Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln 305 310 315 320 atc atg gcc agc ccc gtc ggc ttc agc ggc ccc gag ttc acc ttc ccc 1008 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro                 325 330 335 ctg tac ggc acc atg ggc aac gct gca cct cag cag cgc atc gtg gca 1056 Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala             340 345 350 cag ctg ggc cag gga gtg tac cgc acc ctg agc agc acc ctg tac cgt 1104 Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg         355 360 365 cga cct ttc aac atc ggc atc aac aac cag cag ctg agc gtg ctg gac 1152 Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp     370 375 380 ggc acc gag ttc gcc tac ggc acc agc agc aac ctg ccc agc gcc gtg 1200 Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400 tac cgc aag agc ggc acc gtg gac agc ctg gac gag atc ccc cct cag 1248 Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln                 405 410 415 aac aac aac gtg cca cct cga cag ggc ttc agc cac cgt ctg agc cac 1296 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His             420 425 430 gtg agc atg ttc cgc agt ggc ttc agc aac agc agc gtg agc atc atc 1344 Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile         435 440 445 cgt gca ccc atg ttc agc tgg att cac cgc agc gcc acc ctg acc aac 1392 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Thr Leu Thr Asn     450 455 460 acc atc gac ccc gag cgc atc aac cag atc ccc ctg gtg aag ggc ttc 1440 Thr Ile Asp Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe 465 470 475 480 cgg gtg tgg ggc ggc acc agc gtg atc acc ggc ccc ggc ttc acc gga 1488 Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly                 485 490 495 ggc gac atc ctg cgc aga aac acc ttc ggc gac ttc gtg agc ctg cag 1536 Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln             500 505 510 gtg aac atc aac agc ccc atc acc cag cgt tac cgc ctg cgc ttc cgc 1584 Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg         515 520 525 tac gcc agc agc cgc gac gcc cgt gtg atc gtg ctg act ggc gcc gct 1632 Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala     530 535 540 agc acc ggt gtg ggc ggt cag gtg agc gtg aac atg ccc ctg cag aag 1680 Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys 545 550 555 560 act atg gag atc ggc gag aac ctg act agt cgc acc ttc cgc tac acc 1728 Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr                 565 570 575 gac ttc agc aac ccc ttc agc ttc cgc gcc aac ccc gac atc atc ggc 1776 Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly             580 585 590 atc agc gag cag ccc ctg ttc ggt gcc ggc agc atc agc agc ggc gag 1824 Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu         595 600 605 ctg tac atc gac aag atc gag atc atc ctg gcc gac gcc acc ttc gag 1872 Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu     610 615 620 gcc gag agc gac ctg gag cgc gcc cag aag gcc gtg aac gcc ctg ttc 1920 Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Ala Leu Phe 625 630 635 640 acc agc agc aac cag atc ggc ctg aag acc gac gtg acc gac tac cac 1968 Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His                 645 650 655 atc gac cag gtg agc aac ctg gtg gac tgc tta agc gac gag ttc tgc 2016 Ile Asp Gln Val Ser Asn Leu Val Asp Cys Leu Ser Asp Glu Phe Cys             660 665 670 ctg gac gag aag aag gag ctg agc gag aag gtg aag cac gcc aag cgc 2064 Leu Asp Glu Lys Lys Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg         675 680 685 ctg agc gac gag cgc aac ctg ctg cag gac ccc aac ttc cgc ggc atc 2112 Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe Arg Gly Ile     690 695 700 aac cgc cag ctg gac cgc ggc tgg cga ggc agc acc gat atc acc atc 2160 Asn Arg Gln Leu Asp Arg Gly Trp Arg Gly Ser Thr Asp Ile Thr Ile 705 710 715 720 cag ggc ggc gac gac gtg ttc aag gag aac tac gtg acc ctg cag ggc 2208 Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Gln Gly                 725 730 735 acc ttc gac gag tgc tac ccc acc tac ctg tac cag ccg atc gac gag 2256 Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Pro Ile Asp Glu             740 745 750 agc aag ctg aag gcc tac acc cgc tac cag ctg cgc ggc tac atc gag 2304 Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg Gly Tyr Ile Glu         755 760 765 gac agc cag gac ctg gaa atc tac ctg atc cgc tac aac gcg aag cac 2352 Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His     770 775 780 gag acc gtg aac gtg ccc ggc acc ggc agc ctg tgg ccc ctg agc gcc 2400 Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Leu Ser Ala 785 790 795 800 ccc agc ccc atc ggc aag tgc ggg gag ccg aat cga tgc gct ccg cac 2448 Pro Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn Arg Cys Ala Pro His                 805 810 815 ctg gag tgg aac ccg gac cta gac tgc agc tgc agg gac ggg gag aag 2496 Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg Asp Gly Glu Lys             820 825 830 tgc gcc cac cac agc cac cac ttc agc ctg gac atc gac gtg ggc tgc 2544 Cys Ala His His Ser His His Phe Ser Leu Asp Ile Asp Val Gly Cys         835 840 845 acc gac ctg aac gag gac ctg ggc gtg tgg gtg atc ttc aag atc aag 2592 Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe Lys Ile Lys     850 855 860 acc cag gac ggc cac gcc cgc ctg ggc aat cta gag ttc ctg gag gag 2640 Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu 865 870 875 880 aag ccc ctg gtg ggc gag gcc ctg gcc cgc gtg aag cgt gct gag aag 2688 Lys Pro Leu Val Gly Glu Ala Leu Ala Arg Val Lys Arg Ala Glu Lys                 885 890 895 aag tgg cgc gac aag cgc gag aag ctg gag tgg gag acc aac atc gtg 2736 Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu Trp Glu Thr Asn Ile Val             900 905 910 tac aag gag gcc aag gag agc gtg gac gcc ctg ttc gtg aac agc cag 2784 Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val Asn Ser Gln         915 920 925 tac gac cgc ctg cag gcc gac acc aac atc gcc atg atc cac gcc gcc 2832 Tyr Asp Arg Leu Gln Ala Asp Thr Asn Ile Ala Met Ile His Ala Ala     930 935 940 gac aag cgc gtg cac agc att cgc gag gcc tac ctg ccc gag ctg agc 2880 Asp Lys Arg Val His Ser Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser 945 950 955 960 gtg atc ccc ggt gtg aac gcc gcc atc ttc gag gaa ctc gag ggc cgc 2928 Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu Glu Gly Arg                 965 970 975 atc ttc acc gcc ttc agc ctg tac gac gcc cgc aac gtg atc aag aac 2976 Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala Arg Asn Val Ile Lys Asn             980 985 990 ggc gac ttc aac aac ggc ctg agc tgc tgg aac gtg aag ggc cac gtg 3024 Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp Asn Val Lys Gly His Val         995 1000 1005 gac gtg gag gag cag aac aac cac cgc agc gtg ctg gtg gtg ccc gag 3072 Asp Val Glu Glu Gln Asn Asn His Arg Ser Val Leu Val Val Pro Glu    1010 1015 1020 tgg gag gcc gag gtg agc cag gag gtg cgc gtg tgc ccc ggc cgc ggc 3120 Trp Glu Ala Glu Val Ser Gln Glu Val Arg Val Cys Pro Gly Arg Gly 1025 1030 1035 1040 tac atc ctg cgc gtg acc gcc tac aag gag ggc tac ggc gag ggc tgc 3168 Tyr Ile Leu Arg Val Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys                1045 1050 1055 gtg acc atc cac gag atc gag aac aac acc gac gag ctc aag ttc agc 3216 Val Thr Ile His Glu Ile Glu Asn Asn Thr Asp Glu Leu Lys Phe Ser            1060 1065 1070 aac tgc gtg gag gag gag gtt tac ccc aac aac acc gtg acc tgc aac 3264 Asn Cys Val Glu Glu Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn        1075 1080 1085 gac tac acc gcg acc cag gag gag tac gaa ggc acc tac acc tct cgc 3312 Asp Tyr Thr Ala Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg    1090 1095 1100 aac agg ggt tac gac ggc gcc tac gag tcc aac agc tcc gtg cca gct 3360 Asn Arg Gly Tyr Asp Gly Ala Tyr Glu Ser Asn Ser Ser Val Pro Ala 1105 1110 1115 1120 gac tac gcc agc gcc tac gag gag aaa gcc tac acc gac ggt aga cgc 3408 Asp Tyr Ala Ser Ala Tyr Glu Glu Lys Ala Tyr Thr Asp Gly Arg Arg                1125 1130 1135 gac aac cca tgt gag agc aac aga ggc tac ggc gac tac acc ccc ctg 3456 Asp Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu            1140 1145 1150 ccc gct gga tac gtg acc aag gag ctg gag tac ttc ccc gag acc gac 3504 Pro Ala Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr Asp        1155 1160 1165 aag gtg tgg atc gag att ggc gag acc gag ggc acc ttc atc gtg gac 3552 Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val Asp    1170 1175 1180 agc gtg gag ctg ctg ctg atg gag gag tag 3582 Ser Val Glu Leu Leu Leu Met Glu Glu 1185 1190 <210> 8 <211> 1193 <212> PRT <213> Artificial Sequence <223> Description of Artificial Sequence: synthetic gene       encoding H04 with full-length Cry1Ab tail <400> 8 Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu   1 5 10 15 Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly              20 25 30 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser          35 40 45 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile      50 55 60 Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile  65 70 75 80 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala                  85 90 95 Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu             100 105 110 Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu         115 120 125 Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala     130 135 140 Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155 160 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser                 165 170 175 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg             180 185 190 Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val         195 200 205 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg     210 215 220 Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro                 245 250 255 Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val             260 265 270 Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu         275 280 285 Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr     290 295 300 Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln 305 310 315 320 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro                 325 330 335 Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala             340 345 350 Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg         355 360 365 Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp     370 375 380 Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400 Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln                 405 410 415 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His             420 425 430 Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile         435 440 445 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Thr Leu Thr Asn     450 455 460 Thr Ile Asp Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe 465 470 475 480 Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly                 485 490 495 Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln             500 505 510 Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg         515 520 525 Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala     530 535 540 Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys 545 550 555 560 Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr                 565 570 575 Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly             580 585 590 Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu         595 600 605 Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu     610 615 620 Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Ala Leu Phe 625 630 635 640 Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His                 645 650 655 Ile Asp Gln Val Ser Asn Leu Val Asp Cys Leu Ser Asp Glu Phe Cys             660 665 670 Leu Asp Glu Lys Lys Glu Leu Ser Glu Lys Val Lys His Ala Lys Arg         675 680 685 Leu Ser Asp Glu Arg Asn Leu Leu Gln Asp Pro Asn Phe Arg Gly Ile     690 695 700 Asn Arg Gln Leu Asp Arg Gly Trp Arg Gly Ser Thr Asp Ile Thr Ile 705 710 715 720 Gln Gly Gly Asp Asp Val Phe Lys Glu Asn Tyr Val Thr Leu Gln Gly                 725 730 735 Thr Phe Asp Glu Cys Tyr Pro Thr Tyr Leu Tyr Gln Pro Ile Asp Glu             740 745 750 Ser Lys Leu Lys Ala Tyr Thr Arg Tyr Gln Leu Arg Gly Tyr Ile Glu         755 760 765 Asp Ser Gln Asp Leu Glu Ile Tyr Leu Ile Arg Tyr Asn Ala Lys His     770 775 780 Glu Thr Val Asn Val Pro Gly Thr Gly Ser Leu Trp Pro Leu Ser Ala 785 790 795 800 Pro Ser Pro Ile Gly Lys Cys Gly Glu Pro Asn Arg Cys Ala Pro His                 805 810 815 Leu Glu Trp Asn Pro Asp Leu Asp Cys Ser Cys Arg Asp Gly Glu Lys             820 825 830 Cys Ala His His Ser His His Phe Ser Leu Asp Ile Asp Val Gly Cys         835 840 845 Thr Asp Leu Asn Glu Asp Leu Gly Val Trp Val Ile Phe Lys Ile Lys     850 855 860 Thr Gln Asp Gly His Ala Arg Leu Gly Asn Leu Glu Phe Leu Glu Glu 865 870 875 880 Lys Pro Leu Val Gly Glu Ala Leu Ala Arg Val Lys Arg Ala Glu Lys                 885 890 895 Lys Trp Arg Asp Lys Arg Glu Lys Leu Glu Trp Glu Thr Asn Ile Val             900 905 910 Tyr Lys Glu Ala Lys Glu Ser Val Asp Ala Leu Phe Val Asn Ser Gln         915 920 925 Tyr Asp Arg Leu Gln Ala Asp Thr Asn Ile Ala Met Ile His Ala Ala     930 935 940 Asp Lys Arg Val His Ser Ile Arg Glu Ala Tyr Leu Pro Glu Leu Ser 945 950 955 960 Val Ile Pro Gly Val Asn Ala Ala Ile Phe Glu Glu Leu Glu Gly Arg                 965 970 975 Ile Phe Thr Ala Phe Ser Leu Tyr Asp Ala Arg Asn Val Ile Lys Asn             980 985 990 Gly Asp Phe Asn Asn Gly Leu Ser Cys Trp Asn Val Lys Gly His Val         995 1000 1005 Asp Val Glu Glu Gln Asn Asn His Arg Ser Val Leu Val Val Pro Glu    1010 1015 1020 Trp Glu Ala Glu Val Ser Gln Glu Val Arg Val Cys Pro Gly Arg Gly 1025 1030 1035 1040 Tyr Ile Leu Arg Val Thr Ala Tyr Lys Glu Gly Tyr Gly Glu Gly Cys                1045 1050 1055 Val Thr Ile His Glu Ile Glu Asn Asn Thr Asp Glu Leu Lys Phe Ser            1060 1065 1070 Asn Cys Val Glu Glu Glu Val Tyr Pro Asn Asn Thr Val Thr Cys Asn        1075 1080 1085 Asp Tyr Thr Ala Thr Gln Glu Glu Tyr Glu Gly Thr Tyr Thr Ser Arg    1090 1095 1100 Asn Arg Gly Tyr Asp Gly Ala Tyr Glu Ser Asn Ser Ser Val Pro Ala 1105 1110 1115 1120 Asp Tyr Ala Ser Ala Tyr Glu Glu Lys Ala Tyr Thr Asp Gly Arg Arg                1125 1130 1135 Asp Asn Pro Cys Glu Ser Asn Arg Gly Tyr Gly Asp Tyr Thr Pro Leu            1140 1145 1150 Pro Ala Gly Tyr Val Thr Lys Glu Leu Glu Tyr Phe Pro Glu Thr Asp        1155 1160 1165 Lys Val Trp Ile Glu Ile Gly Glu Thr Glu Gly Thr Phe Ile Val Asp    1170 1175 1180 Ser Val Glu Leu Leu Leu Met Glu Glu 1185 1190 <210> 9 <211> 2007 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: synthetic gene       encoding H04 plus the first 40 amino acids of the       Cry1Ab tail <220> <221> CDS (222) (1) .. (2007) <223> H04 with truncated Cry1Ab tail <400> 9 atg gac aac aac ccc aac atc aac gag tgc atc ccc tac aac tgc ctg 48 Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu   1 5 10 15 agc aac ccc gag gtg gag gtg ctg ggc ggc gag cgc atc gag acc ggc 96 Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly              20 25 30 tac acc ccc atc gac atc agc ctg agc ctg acc cag ttc ctg ctg agc 144 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser          35 40 45 gag ttc gtg ccc ggc gcc ggc ttc gtg ctg ggc ctg gtg gac atc atc 192 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile      50 55 60 tgg ggc atc ttc ggc ccc agc cag tgg gac gcc ttc ctg gtg cag atc 240 Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile  65 70 75 80 gag cag ttg ata aac caa cgc ata gag gaa ttc gcc cgc aac cag gcc 288 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala                  85 90 95 atc agc cgc ctg gag ggc ctg agc aac ctg tac caa atc tac gcc gag 336 Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu             100 105 110 agc ttc cgc gag tgg gag gcc gac ccc acc aac ccc gcc ctg cgc gag 384 Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu         115 120 125 gag atg cgc atc cag ttc aac gac atg aac agc gcc ctg acc acc gcc 432 Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala     130 135 140 atc ccc ctg ttc gcc gtg cag aac tac cag gtg ccc ctg ctg agc gtg 480 Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155 160 tac gtg cag gcc gcc aac ctg cac ctg agc gtg ctg cgc gac gtc agc 528 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser                 165 170 175 gtg ttc ggc cag cgc tgg ggc ttc gac gcc gcc acc atc aac agc cgc 576 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg             180 185 190 tac aac gac ctg acc cgc ctg atc ggc aac tac acc gac cac gcc gtg 624 Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val         195 200 205 cgc tgg tac aac acc ggc ctg gag cgc gtg tgg ggt ccc gac agc cgc 672 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg     210 215 220 gac tgg atc agg tac aac cag ttc cgc cgc gag ctg acc ctg acc gtg 720 Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 ctg gac atc gtg agc ctg ttc ccc aac tac gac agc cgc acc tac ccc 768 Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro                 245 250 255 atc cgc acc gtg agc cag ctg acc cgc gag att tac acc aac ccc gtg 816 Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val             260 265 270 ctg gag aac ttc gac ggc agc ttc cgc ggc agc gcc cag ggc atc gag 864 Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu         275 280 285 ggc agc atc cgc agc ccc cac ctg atg gac atc ctg aac agc atc acc 912 Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr     290 295 300 atc tac acc gac gcc cac cgc ggc gag tac tac tgg agc ggc cac cag 960 Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln 305 310 315 320 atc atg gcc agc ccc gtc ggc ttc agc ggc ccc gag ttc acc ttc ccc 1008 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro                 325 330 335 ctg tac ggc acc atg ggc aac gct gca cct cag cag cgc atc gtg gca 1056 Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala             340 345 350 cag ctg ggc cag gga gtg tac cgc acc ctg agc agc acc ctg tac cgt 1104 Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg         355 360 365 cga cct ttc aac atc ggc atc aac aac cag cag ctg agc gtg ctg gac 1152 Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp     370 375 380 ggc acc gag ttc gcc tac ggc acc agc agc aac ctg ccc agc gcc gtg 1200 Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400 tac cgc aag agc ggc acc gtg gac agc ctg gac gag atc ccc cct cag 1248 Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln                 405 410 415 aac aac aac gtg cca cct cga cag ggc ttc agc cac cgt ctg agc cac 1296 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His             420 425 430 gtg agc atg ttc cgc agt ggc ttc agc aac agc agc gtg agc atc atc 1344 Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile         435 440 445 cgt gca ccc atg ttc agc tgg att cac cgc agc gcc acc ctg acc aac 1392 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Thr Leu Thr Asn     450 455 460 acc atc gac ccc gag cgc atc aac cag atc ccc ctg gtg aag ggc ttc 1440 Thr Ile Asp Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe 465 470 475 480 cgg gtg tgg ggc ggc acc agc gtg atc acc ggc ccc ggc ttc acc gga 1488 Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly                 485 490 495 ggc gac atc ctg cgc aga aac acc ttc ggc gac ttc gtg agc ctg cag 1536 Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln             500 505 510 gtg aac atc aac agc ccc atc acc cag cgt tac cgc ctg cgc ttc cgc 1584 Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg         515 520 525 tac gcc agc agc cgc gac gcc cgt gtg atc gtg ctg act ggc gcc gct 1632 Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala     530 535 540 agc acc ggt gtg ggc ggt cag gtg agc gtg aac atg ccc ctg cag aag 1680 Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys 545 550 555 560 act atg gag atc ggc gag aac ctg act agt cgc acc ttc cgc tac acc 1728 Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr                 565 570 575 gac ttc agc aac ccc ttc agc ttc cgc gcc aac ccc gac atc atc ggc 1776 Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly             580 585 590 atc agc gag cag ccc ctg ttc ggt gcc ggc agc atc agc agc ggc gag 1824 Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu         595 600 605 ctg tac atc gac aag atc gag atc atc ctg gcc gac gcc acc ttc gag 1872 Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu     610 615 620 gcc gag agc gac ctg gag cgc gcc cag aag gcc gtg aac gcc ctg ttc 1920 Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Ala Leu Phe 625 630 635 640 acc agc agc aac cag atc ggc ctg aag acc gac gtg acc gac tac cac 1968 Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His                 645 650 655 atc gac cag gtg agc aac ctg gtg gac tgc tta agc tag 2007 Ile Asp Gln Val Ser Asn Leu Val Asp Cys Leu Ser             660 665 <210> 10 <211> 668 <212> PRT <213> Artificial Sequence <223> Description of Artificial Sequence: synthetic gene       encoding H04 plus the first 40 amino acids of the       Cry1Ab tail <400> 10 Met Asp Asn Asn Pro Asn Ile Asn Glu Cys Ile Pro Tyr Asn Cys Leu   1 5 10 15 Ser Asn Pro Glu Val Glu Val Leu Gly Gly Glu Arg Ile Glu Thr Gly              20 25 30 Tyr Thr Pro Ile Asp Ile Ser Leu Ser Leu Thr Gln Phe Leu Leu Ser          35 40 45 Glu Phe Val Pro Gly Ala Gly Phe Val Leu Gly Leu Val Asp Ile Ile      50 55 60 Trp Gly Ile Phe Gly Pro Ser Gln Trp Asp Ala Phe Leu Val Gln Ile  65 70 75 80 Glu Gln Leu Ile Asn Gln Arg Ile Glu Glu Phe Ala Arg Asn Gln Ala                  85 90 95 Ile Ser Arg Leu Glu Gly Leu Ser Asn Leu Tyr Gln Ile Tyr Ala Glu             100 105 110 Ser Phe Arg Glu Trp Glu Ala Asp Pro Thr Asn Pro Ala Leu Arg Glu         115 120 125 Glu Met Arg Ile Gln Phe Asn Asp Met Asn Ser Ala Leu Thr Thr Ala     130 135 140 Ile Pro Leu Phe Ala Val Gln Asn Tyr Gln Val Pro Leu Leu Ser Val 145 150 155 160 Tyr Val Gln Ala Ala Asn Leu His Leu Ser Val Leu Arg Asp Val Ser                 165 170 175 Val Phe Gly Gln Arg Trp Gly Phe Asp Ala Ala Thr Ile Asn Ser Arg             180 185 190 Tyr Asn Asp Leu Thr Arg Leu Ile Gly Asn Tyr Thr Asp His Ala Val         195 200 205 Arg Trp Tyr Asn Thr Gly Leu Glu Arg Val Trp Gly Pro Asp Ser Arg     210 215 220 Asp Trp Ile Arg Tyr Asn Gln Phe Arg Arg Glu Leu Thr Leu Thr Val 225 230 235 240 Leu Asp Ile Val Ser Leu Phe Pro Asn Tyr Asp Ser Arg Thr Tyr Pro                 245 250 255 Ile Arg Thr Val Ser Gln Leu Thr Arg Glu Ile Tyr Thr Asn Pro Val             260 265 270 Leu Glu Asn Phe Asp Gly Ser Phe Arg Gly Ser Ala Gln Gly Ile Glu         275 280 285 Gly Ser Ile Arg Ser Pro His Leu Met Asp Ile Leu Asn Ser Ile Thr     290 295 300 Ile Tyr Thr Asp Ala His Arg Gly Glu Tyr Tyr Trp Ser Gly His Gln 305 310 315 320 Ile Met Ala Ser Pro Val Gly Phe Ser Gly Pro Glu Phe Thr Phe Pro                 325 330 335 Leu Tyr Gly Thr Met Gly Asn Ala Ala Pro Gln Gln Arg Ile Val Ala             340 345 350 Gln Leu Gly Gln Gly Val Tyr Arg Thr Leu Ser Ser Thr Leu Tyr Arg         355 360 365 Arg Pro Phe Asn Ile Gly Ile Asn Asn Gln Gln Leu Ser Val Leu Asp     370 375 380 Gly Thr Glu Phe Ala Tyr Gly Thr Ser Ser Asn Leu Pro Ser Ala Val 385 390 395 400 Tyr Arg Lys Ser Gly Thr Val Asp Ser Leu Asp Glu Ile Pro Pro Gln                 405 410 415 Asn Asn Asn Val Pro Pro Arg Gln Gly Phe Ser His Arg Leu Ser His             420 425 430 Val Ser Met Phe Arg Ser Gly Phe Ser Asn Ser Ser Val Ser Ile Ile         435 440 445 Arg Ala Pro Met Phe Ser Trp Ile His Arg Ser Ala Thr Leu Thr Asn     450 455 460 Thr Ile Asp Pro Glu Arg Ile Asn Gln Ile Pro Leu Val Lys Gly Phe 465 470 475 480 Arg Val Trp Gly Gly Thr Ser Val Ile Thr Gly Pro Gly Phe Thr Gly                 485 490 495 Gly Asp Ile Leu Arg Arg Asn Thr Phe Gly Asp Phe Val Ser Leu Gln             500 505 510 Val Asn Ile Asn Ser Pro Ile Thr Gln Arg Tyr Arg Leu Arg Phe Arg         515 520 525 Tyr Ala Ser Ser Arg Asp Ala Arg Val Ile Val Leu Thr Gly Ala Ala     530 535 540 Ser Thr Gly Val Gly Gly Gln Val Ser Val Asn Met Pro Leu Gln Lys 545 550 555 560 Thr Met Glu Ile Gly Glu Asn Leu Thr Ser Arg Thr Phe Arg Tyr Thr                 565 570 575 Asp Phe Ser Asn Pro Phe Ser Phe Arg Ala Asn Pro Asp Ile Ile Gly             580 585 590 Ile Ser Glu Gln Pro Leu Phe Gly Ala Gly Ser Ile Ser Ser Gly Glu         595 600 605 Leu Tyr Ile Asp Lys Ile Glu Ile Ile Leu Ala Asp Ala Thr Phe Glu     610 615 620 Ala Glu Ser Asp Leu Glu Arg Ala Gln Lys Ala Val Asn Ala Leu Phe 625 630 635 640 Thr Ser Ser Asn Gln Ile Gly Leu Lys Thr Asp Val Thr Asp Tyr His                 645 650 655 Ile Asp Gln Val Ser Asn Leu Val Asp Cys Leu Ser             660 665 <210> 11 <211> 13269 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: pNOV1308 <220> <221> misc_feature (222) (1) .. (1896) <223> synthetic nucleotide sequence encoding the toxin       portion of H04, without a tail <220> <221> misc_feature (2102) .. (4083) <223> Zea mays ubiquitin promoter <220> <221> misc_feature (222) (4180) .. (5283) <223> PMI marker gene <220> <221> misc_feature (222) (11247) .. (12647) <223> Zm Ubi promoter <400> 11 atggacaaca accccaacat caacgagtgc atcccctaca actgcctgag caaccccgag 60 gtggaggtgc tgggcggcga gcgcatcgag accggctaca cccccatcga catcagcctg 120 agcctgaccc agttcctgct gagcgagttc gtgcccggcg ccggcttcgt gctgggcctg 180 gtggacatca tctggggcat cttcggcccc agccagtggg acgccttcct ggtgcagatc 240 gagcagttga taaaccaacg catagaggaa ttcgcccgca accaggccat cagccgcctg 300 gagggcctga gcaacctgta ccaaatctac gccgagagct tccgcgagtg ggaggccgac 360 cccaccaacc ccgccctgcg cgaggagatg cgcatccagt tcaacgacat gaacagcgcc 420 ctgaccaccg ccatccccct gttcgccgtg cagaactacc aggtgcccct gctgagcgtg 480 tacgtgcagg ccgccaacct gcacctgagc gtgctgcgcg acgtcagcgt gttcggccag 540 cgctggggct tcgacgccgc caccatcaac agccgctaca acgacctgac ccgcctgatc 600 ggcaactaca ccgaccacgc cgtgcgctgg tacaacaccg gcctggagcg cgtgtggggt 660 cccgacagcc gcgactggat caggtacaac cagttccgcc gcgagctgac cctgaccgtg 720 ctggacatcg tgagcctgtt ccccaactac gacagccgca cctaccccat ccgcaccgtg 780 agccagctga cccgcgagat ttacaccaac cccgtgctgg agaacttcga cggcagcttc 840 cgcggcagcg cccagggcat cgagggcagc atccgcagcc cccacctgat ggacatcctg 900 aacagcatca ccatctacac cgacgcccac cgcggcgagt actactggag cggccaccag 960 atcatggcca gccccgtcgg cttcagcggc cccgagttca ccttccccct gtacggcacc 1020 atgggcaacg ctgcacctca gcagcgcatc gtggcacagc tgggccaggg agtgtaccgc 1080 accctgagca gcaccctgta ccgtcgacct ttcaacatcg gcatcaacaa ccagcagctg 1140 agcgtgctgg acggcaccga gttcgcctac ggcaccagca gcaacctgcc cagcgccgtg 1200 taccgcaaga gcggcaccgt ggacagcctg gacgagatcc cccctcagaa caacaacgtg 1260 ccacctcgac agggcttcag ccaccgtctg agccacgtga gcatgttccg cagtggcttc 1320 agcaacagca gcgtgagcat catccgtgca cccatgttca gctggattca ccgcagcgcc 1380 accctgacca acaccatcga ccccgagcgc atcaaccaga tccccctggt gaagggcttc 1440 cgggtgtggg gcggcaccag cgtgatcacc ggccccggct tcaccggagg cgacatcctg 1500 cgcagaaaca ccttcggcga cttcgtgagc ctgcaggtga acatcaacag ccccatcacc 1560 cagcgttacc gcctgcgctt ccgctacgcc agcagccgcg acgcccgtgt gatcgtgctg 1620 actggcgccg ctagcaccgg tgtgggcggt caggtgagcg tgaacatgcc cctgcagaag 1680 actatggaga tcggcgagaa cctgactagt cgcaccttcc gctacaccga cttcagcaac 1740 cccttcagct tccgcgccaa ccccgacatc atcggcatca gcgagcagcc cctgttcggt 1800 gccggcagca tcagcagcgg cgagctgtac atcgacaaga tcgagatcat cctggccgac 1860 gccaccttcg aggccgagag cgacctggag cgctaagatc tgttctgcac aaagtggagt 1920 agtcagtcat cgatcaggaa ccagacacca gacttttatt catacagtga agtgaagtga 1980 agtgcagtgc agtgagttgc tggtttttgt acaacttagt atgtatttgt atttgtaaaa 2040 tacttctatc aataaaattt ctaattccta aaaccaaaat ccaggggtac cagcttgcat 2100 gcctgcagtg cagcgtgacc cggtcgtgcc cctctctaga gataatgagc attgcatgtc 2160 taagttataa aaaattacca catatttttt ttgtcacact tgtttgaagt gcagtttatc 2220 tatctttata catatattta aactttactc tacgaataat ataatctata gtactacaat 2280 aatatcagtg ttttagagaa tcatataaat gaacagttag acatggtcta aaggacaatt 2340 gagtattttg acaacaggac tctacagttt tatcttttta gtgtgcatgt gttctccttt 2400 ttttttgcaa atagcttcac ctatataata cttcatccat tttattagta catccattta 2460 gggtttaggg ttaatggttt ttatagacta atttttttag tacatctatt ttattctatt 2520 ttagcctcta aattaagaaa actaaaactc tattttagtt tttttattta ataatttaga 2580 tataaaatag aataaaataa agtgactaaa aattaaacaa atacccttta agaaattaaa 2640 aaaactaagg aaacattttt cttgtttcga gtagataatg ccagcctgtt aaacgccgtc 2700 gacgagtcta acggacacca accagcgaac cagcagcgtc gcgtcgggcc aagcgaagca 2760 gacggcacgg catctctgtc gctgcctctg gacccctctc gagagttccg ctccaccgtt 2820 ggacttgctc cgctgtcggc atccagaaat tgcgtggcgg agcggcagac gtgagccggc 2880 acggcaggcg gcctcctcct cctctcacgg caccggcagc tacgggggat tcctttccca 2940 ccgctccttc gctttccctt cctcgcccgc cgtaataaat agacaccccc tccacaccct 3000 ctttccccaa cctcgtgttg ttcggagcgc acacacacac aaccagatct cccccaaatc 3060 cacccgtcgg cacctccgct tcaaggtacg ccgctcgtcc tccccccccc cccctctcta 3120 ccttctctag atcggcgttc cggtccatgg ttagggcccg gtagttctac ttctgttcat 3180 gtttgtgtta gatccgtgtt tgtgttagat ccgtgctgct agcgttcgta cacggatgcg 3240 acctgtacgt cagacacgtt ctgattgcta acttgccagt gtttctcttt ggggaatcct 3300 gggatggctc tagccgttcc gcagacggga tcgatttcat gatttttttt gtttcgttgc 3360 atagggtttg gtttgccctt ttcctttatt tcaatatatg ccgtgcactt gtttgtcggg 3420 tcatcttttc atgctttttt ttgtcttggt tgtgatgatg tggtctggtt gggcggtcgt 3480 tctagatcgg agtagaattc tgtttcaaac tacctggtgg atttattaat tttggatctg 3540 tatgtgtgtg ccatacatat tcatagttac gaattgaaga tgatggatgg aaatatcgat 3600 ctaggatagg tatacatgtt gatgcgggtt ttactgatgc atatacagag atgctttttg 3660 ttcgcttggt tgtgatgatg tggtgtggtt gggcggtcgt tcattcgttc tagatcggag 3720 tagaatactg tttcaaacta cctggtgtat ttattaattt tggaactgta tgtgtgtgtc 3780 atacatcttc atagttacga gtttaagatg gatggaaata tcgatctagg ataggtatac 3840 atgttgatgt gggttttact gatgcatata catgatggca tatgcagcat ctattcatat 3900 gctctaacct tgagtaccta tctattataa taaacaagta tgttttataa ttattttgat 3960 cttgatatac ttggatgatg gcatatgcag cagctatatg tggatttttt tagccctgcc 4020 ttcatacgct atttatttgc ttggtactgt ttcttttgtc gatgctcacc ctgttgtttg 4080 gtgttacttc tgcagggatc cccgatcatg caaaaactca ttaactcagt gcaaaactat 4140 gcctggggca gcaaaacggc gttgactgaa ctttatggta tggaaaatcc gtccagccag 4200 ccgatggccg agctgtggat gggcgcacat ccgaaaagca gttcacgagt gcagaatgcc 4260 gccggagata tcgtttcact gcgtgatgtg attgagagtg ataaatcgac tctgctcgga 4320 gaggccgttg ccaaacgctt tggcgaactg cctttcctgt tcaaagtatt atgcgcagca 4380 cagccactct ccattcaggt tcatccaaac aaacacaatt ctgaaatcgg ttttgccaaa 4440 gaaaatgccg caggtatccc gatggatgcc gccgagcgta actataaaga tcctaaccac 4500 aagccggagc tggtttttgc gctgacgcct ttccttgcga tgaacgcgtt tcgtgaattt 4560 tccgagattg tctccctact ccagccggtc gcaggtgcac atccggcgat tgctcacttt 4620 ttacaacagc ctgatgccga acgtttaagc gaactgttcg ccagcctgtt gaatatgcag 4680 ggtgaagaaa aatcccgcgc gctggcgatt ttaaaatcgg ccctcgatag ccagcagggt 4740 gaaccgtggc aaacgattcg tttaatttct gaattttacc cggaagacag cggtctgttc 4800 tccccgctat tgctgaatgt ggtgaaattg aaccctggcg aagcgatgtt cctgttcgct 4860 gaaacaccgc acgcttacct gcaaggcgtg gcgctggaag tgatggcaaa ctccgataac 4920 gtgctgcgtg cgggtctgac gcctaaatac attgatattc cggaactggt tgccaatgtg 4980 aaattcgaag ccaaaccggc taaccagttg ttgacccagc cggtgaaaca aggtgcagaa 5040 ctggacttcc cgattccagt ggatgatttt gccttctcgc tgcatgacct tagtgataaa 5100 gaaaccacca ttagccagca gagtgccgcc attttgttct gcgtcgaagg cgatgcaacg 5160 ttgtggaaag gttctcagca gttacagctt aaaccgggtg aatcagcgtt tattgccgcc 5220 aacgaatcac cggtgactgt caaaggccac ggccgtttag cgcgtgttta caacaagctg 5280 taagagctta ctgaaaaaat taacatctct tgctaagctg ggagctcgat ccgtcgacct 5340 gcagatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt 5400 gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa 5460 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa 5520 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca 5580 tctatgttac tagatctgct agccctgcag gaaatttacc ggtgcccggg cggccagcat 5640 ggccgtatcc gcaatgtgtt attaagttgt ctaagcgtca atttgtttac accacaatat 5700 atcctgccac cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc 5760 gatacaggca gcccatcaga attaattctc atgtttgaca gcttatcatc gactgcacgg 5820 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg 5880 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg 5940 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc 6000 cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga aacagaccat 6060 gagggaagcg ttgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga 6120 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg 6180 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac 6240 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga 6300 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta 6360 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat 6420 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca 6480 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga 6540 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg 6600 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa 6660 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca 6720 gcccgtcata cttgaagcta ggcaggctta tcttggacaa gaagatcgct tggcctcgcg 6780 cgcagatcag ttggaagaat ttgttcacta cgtgaaaggc gagatcacca aagtagtcgg 6840 caaataaagc tctagtggat ctccgtaccc ccgggggatc tggctcgcgg cggacgcacg 6900 acgccggggc gagaccatag gcgatctcct aaatcaatag tagctgtaac ctcgaagcgt 6960 ttcacttgta acaacgattg agaatttttg tcataaaatt gaaatacttg gttcgcattt 7020 ttgtcatccg cggtcagccg caattctgac gaactgccca tttagctgga gatgattgta 7080 catccttcac gtgaaaattt ctcaagcgct gtgaacaagg gttcagattt tagattgaaa 7140 ggtgagccgt tgaaacacgt tcttcttgtc gatgacgacg tcgctatgcg gcatcttatt 7200 attgaatacc ttacgatcca cgccttcaaa gtgaccgcgg tagccgacag cacccagttc 7260 acaagagtac tctcttccgc gacggtcgat gtcgtggttg ttgatctaaa tttaggtcgt 7320 gaagatgggc tcgagatcgt tcgtaatctg gcggcaaagt ctgatattcc aatcataatt 7380 atcagtggcg accgccttga ggagacggat aaagttgttg cactcgagct aggagcaagt 7440 gattttatcg ctaagccgtt cagtatcaga gagtttctag cacgcattcg ggttgccttg 7500 cgcgtgcgcc ccaacgttgt ccgctccaaa gaccgacggt ctttttgttt tactgactgg 7560 acacttaatc tcaggcaacg tcgcttgatg tccgaagctg gcggtgaggt gaaacttacg 7620 gcaggtgagt tcaatcttct cctcgcgttt ttagagaaac cccgcgacgt tctatcgcgc 7680 gagcaacttc tcattgccag tcgagtacgc gacgaggagg tttatgacag gagtatagat 7740 gttctcattt tgaggctgcg ccgcaaactt gaggcagatc cgtcaagccc tcaactgata 7800 aaaacagcaa gaggtgccgg ttatttcttt gacgcggacg tgcaggtttc gcacgggggg 7860 acgatggcag cctgagccaa ttcccagatc cccgaggaat cggcgtgagc ggtcgcaaac 7920 catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag aagttgaagg 7980 ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt gaatcgtggc 8040 aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt 8100 cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg atgctctatg 8160 acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc 8220 gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac gtagaggttt 8280 ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg atggcggttt 8340 cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag cccggccgcg 8400 tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat ggcggaaagc 8460 agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt gccatgcagc 8520 gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa gccttgatta 8580 gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag atcgagctag 8640 ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg acggttcacc 8700 ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg gcacgccgcg 8760 ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc agtggcagcg 8820 ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc 8880 cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc 8940 gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc 9000 aaattgccct agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat agcacgtaca 9060 ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac ccaaagccgt 9120 acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa ggcgattttt 9180 ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac 9240 tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct 9300 ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca aaaatggctg 9360 gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc 9420 ggcgctgagg tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc tgaatcgccc 9480 catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg taggtggacc 9540 agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg 9600 tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccgc cgtcccgtca 9660 agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc 9720 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg 9780 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag 9840 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc 9900 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga 9960 gaatggcaaa agctctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 10020 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 10080 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 10140 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 10200 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 10260 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 10320 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 10380 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 10440 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 10500 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 10560 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 10620 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 10680 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 10740 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 10800 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 10860 ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg atccggaatt 10920 aattcctgtg gttggcatgc acatacaaat ggacgaacgg ataaaccttt tcacgccctt 10980 ttaaatatcc gattattcta ataaacgctc ttttctctta ggtttacccg ccaatatatc 11040 ctgtcaaaca ctgatagttt aaactgaagg cgggaaacga caatctgatc atgagcggag 11100 aattaaggga gtcacgttat gacccccgcc gatgacgcgg gacaagccgt tttacgtttg 11160 gaactgacag aaccgcaacg ctgcaggaat tggccgcagc ggccatttaa atcaattggg 11220 cgcgccgaat tcgagctcgg tacaagcttg catgcctgca gtgcagcgtg acccggtcgt 11280 gcccctctct agagataatg agcattgcat gtctaagtta taaaaaatta ccacatattt 11340 tttttgtcac acttgtttga agtgcagttt atctatcttt atacatatat ttaaacttta 11400 ctctacgaat aatataatct atagtactac aataatatca gtgttttaga gaatcatata 11460 aatgaacagt tagacatggt ctaaaggaca attgagtatt ttgacaacag gactctacag 11520 ttttatcttt ttagtgtgca tgtgttctcc tttttttttg caaatagctt cacctatata 11580 atacttcatc cattttatta gtacatccat ttagggttta gggttaatgg tttttataga 11640 ctaatttttt tagtacatct attttattct attttagcct ctaaattaag aaaactaaaa 11700 ctctatttta gtttttttat ttaataattt agatataaaa tagaataaaa taaagtgact 11760 aaaaattaaa caaataccct ttaagaaatt aaaaaaacta aggaaacatt tttcttgttt 11820 cgagtagata atgccagcct gttaaacgcc gtcgacgagt ctaacggaca ccaaccagcg 11880 aaccagcagc gtcgcgtcgg gccaagcgaa gcagacggca cggcatctct gtcgctgcct 11940 ctggacccct ctcgagagtt ccgctccacc gttggacttg ctccgctgtc ggcatccaga 12000 aattgcgtgg cggagcggca gacgtgagcc ggcacggcag gcggcctcct cctcctctca 12060 cggcacggca gctacggggg attcctttcc caccgctcct tcgctttccc ttcctcgccc 12120 gccgtaataa atagacaccc cctccacacc ctctttcccc aacctcgtgt tgttcggagc 12180 gcacacacac acaaccagat ctcccccaaa tccacccgtc ggcacctccg cttcaaggta 12240 cgccgctcgt cctccccccc cccccctctc taccttctct agatcggcgt tccggtccat 12300 ggttagggcc cggtagttct acttctgttc atgtttgtgt tagatccgtg tttgtgttag 12360 atccgtgctg ctagcgttcg tacacggatg cgacctgtac gtcagacacg ttctgattgc 12420 taacttgcca gtgtttctct ttggggaatc ctgggatggc tctagccgtt ccgcagacgg 12480 gatcgatttc atgatttttt ttgtttcgtt gcatagggtt tggtttgccc ttttccttta 12540 tttcaatata tgccgtgcac ttgtttgtcg ggtcatcttt tcatgctttt ttttgtcttg 12600 gttgtgatga tgtggtctgg ttgggcggtc gttctagatc ggagtagaat tctgtttcaa 12660 actacctggt ggatttatta attttggatc tgtatgtgtg tgccatacat attcatagtt 12720 acgaattgaa gatgatggat ggaaatatcg atctaggata ggtatacatg ttgatgcggg 12780 ttttactgat gcatatacag agatgctttt tgttcgcttg gttgtgatga tgtggtgtgg 12840 ttgggcggtc gttcattcgt tctagatcgg agtagaatac tgtttcaaac tacctggtgt 12900 atttattaat tttggaactg tatgtgtgtg tcatacatct tcatagttac gagtttaaga 12960 tggatggaaa tatcgatcta ggataggtat acatgttgat gtgggtttta ctgatgcata 13020 tacatgatgg catatgcagc atctattcat atgctctaac cttgagtacc tatctattat 13080 aataaacaag tatgttttat aattattttg atcttgatat acttggatga tggcatatgc 13140 agcagctata tgtggatttt tttagccctg ccttcatacg ctatttattt gcttggtact 13200 gtttcttttg tcgatgctca ccctgttgtt tggtgttact tctgcaggtc gactctagag 13260 gatccaaca 13269 <210> 12 <211> 16179 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: pNOV1436 <220> <221> misc_feature (222) (1) .. (3582) <223> synthetic nucleotide sequence encoding the toxin       portion of H04 plus a full-length Cry1Ab tail       portion <220> <221> misc_feature Complement ((10390) .. (11598)) <223> PhosphoMannose Isomerase (PMI) marker gene <220> <221> misc_feature Complement ((12718) .. (13608)) <223> Maize ubiquitin (Zm Ubi) promoter <220> <221> misc_feature <222> (13613) .. (16170) <223> MTL promoter <400> 12 atggacaaca accccaacat caacgagtgc atcccctaca actgcctgag caaccccgag 60 gtggaggtgc tgggcggcga gcgcatcgag accggctaca cccccatcga catcagcctg 120 agcctgaccc agttcctgct gagcgagttc gtgcccggcg ccggcttcgt gctgggcctg 180 gtggacatca tctggggcat cttcggcccc agccagtggg acgccttcct ggtgcagatc 240 gagcagttga taaaccaacg catagaggaa ttcgcccgca accaggccat cagccgcctg 300 gagggcctga gcaacctgta ccaaatctac gccgagagct tccgcgagtg ggaggccgac 360 cccaccaacc ccgccctgcg cgaggagatg cgcatccagt tcaacgacat gaacagcgcc 420 ctgaccaccg ccatccccct gttcgccgtg cagaactacc aggtgcccct gctgagcgtg 480 tacgtgcagg ccgccaacct gcacctgagc gtgctgcgcg acgtcagcgt gttcggccag 540 cgctggggct tcgacgccgc caccatcaac agccgctaca acgacctgac ccgcctgatc 600 ggcaactaca ccgaccacgc cgtgcgctgg tacaacaccg gcctggagcg cgtgtggggt 660 cccgacagcc gcgactggat caggtacaac cagttccgcc gcgagctgac cctgaccgtg 720 ctggacatcg tgagcctgtt ccccaactac gacagccgca cctaccccat ccgcaccgtg 780 agccagctga cccgcgagat ttacaccaac cccgtgctgg agaacttcga cggcagcttc 840 cgcggcagcg cccagggcat cgagggcagc atccgcagcc cccacctgat ggacatcctg 900 aacagcatca ccatctacac cgacgcccac cgcggcgagt actactggag cggccaccag 960 atcatggcca gccccgtcgg cttcagcggc cccgagttca ccttccccct gtacggcacc 1020 atgggcaacg ctgcacctca gcagcgcatc gtggcacagc tgggccaggg agtgtaccgc 1080 accctgagca gcaccctgta ccgtcgacct ttcaacatcg gcatcaacaa ccagcagctg 1140 agcgtgctgg acggcaccga gttcgcctac ggcaccagca gcaacctgcc cagcgccgtg 1200 taccgcaaga gcggcaccgt ggacagcctg gacgagatcc cccctcagaa caacaacgtg 1260 ccacctcgac agggcttcag ccaccgtctg agccacgtga gcatgttccg cagtggcttc 1320 agcaacagca gcgtgagcat catccgtgca cccatgttca gctggattca ccgcagcgcc 1380 accctgacca acaccatcga ccccgagcgc atcaaccaga tccccctggt gaagggcttc 1440 cgggtgtggg gcggcaccag cgtgatcacc ggccccggct tcaccggagg cgacatcctg 1500 cgcagaaaca ccttcggcga cttcgtgagc ctgcaggtga acatcaacag ccccatcacc 1560 cagcgttacc gcctgcgctt ccgctacgcc agcagccgcg acgcccgtgt gatcgtgctg 1620 actggcgccg ctagcaccgg tgtgggcggt caggtgagcg tgaacatgcc cctgcagaag 1680 actatggaga tcggcgagaa cctgactagt cgcaccttcc gctacaccga cttcagcaac 1740 cccttcagct tccgcgccaa ccccgacatc atcggcatca gcgagcagcc cctgttcggt 1800 gccggcagca tcagcagcgg cgagctgtac atcgacaaga tcgagatcat cctggccgac 1860 gccaccttcg aggccgagag cgacctggag cgcgcccaga aggccgtgaa cgccctgttc 1920 accagcagca accagatcgg cctgaagacc gacgtgaccg actaccacat cgaccaggtg 1980 agcaacctgg tggactgctt aagcgacgag ttctgcctgg acgagaagaa ggagctgagc 2040 gagaaggtga agcacgccaa gcgcctgagc gacgagcgca acctgctgca ggaccccaac 2100 ttccgcggca tcaaccgcca gctggaccgc ggctggcgag gcagcaccga tatcaccatc 2160 cagggcggcg acgacgtgtt caaggagaac tacgtgaccc tgcagggcac cttcgacgag 2220 tgctacccca cctacctgta ccagccgatc gacgagagca agctgaaggc ctacacccgc 2280 taccagctgc gcggctacat cgaggacagc caggacctgg aaatctacct gatccgctac 2340 aacgcgaagc acgagaccgt gaacgtgccc ggcaccggca gcctgtggcc cccgagcgcc 2400 cccagcccca tcggcaagtg cggggagccg aatcgatgcg ctccgcacct ggagtggaac 2460 ccggacctag actgcagctg cagggacggg gagaagtgcg cccaccacag ccaccacttc 2520 agcctggaca tcgacgtggg ctgcaccgac ctgaacgagg acctgggcgt gtgggtgatc 2580 ttcaagatca agacccagga cggccacgcc cgcctgggca atctagagtt cctggaggag 2640 aagcccctgg tgggcgaggc cctggcccgc gtgaagcgtg ctgagaagaa gtggcgcgac 2700 aagcgcgaga agctggagtg ggagaccaac atcgtgtaca aggaggccaa ggagagcgtg 2760 gacgccctgt tcgtgaacag ccagtacgac cgcctgcagg ccgacaccaa catcgccatg 2820 atccacgccg ccgacaagcg cgtgcacagc attcgcgagg cctacctgcc cgagctgagc 2880 gtgatccccg gtgtgaacgc cgccatcttc gaggaactcg agggccgcat cttcaccgcc 2940 ttcagcctgt acgacgcccg caacgtgatc aagaacggcg acttcaacaa cggcctgagc 3000 tgctggaacg tgaagggcca cgtggacgtg gaggagcaga acaaccaccg cagcgtgctg 3060 gtggtgcccg agtgggaggc cgaggtgagc caggaggtgc gcgtgtgccc cggccgcggc 3120 tacatcctgc gcgtgaccgc ctacaaggag ggctacggcg agggctgcgt gaccatccac 3180 gagatcgaga acaacaccga cgagctcaag ttcagcaact gcgtggagga ggaggtttac 3240 cccaacaaca ccgtgacctg caacgactac accgcgaccc aggaggagta cgaaggcacc 3300 tacacctctc gcaacagggg ttacgacggc gcctacgagt ccaacagctc cgtgccagct 3360 gactacgcca gcgcccacga ggagaaagcc tacaccgacg gtagacgcga caacccatgt 3420 gagagcaaca gaggctacgg cgactacacc cccctgcccg ctggatacgt gaccaaggag 3480 ctggagtact tccccgagac cgacaaggtg tggatcgaga ttggcgagac cgagggcacc 3540 ttcatcgtgg acagcgtgga gctgctgctg atggaggagt agtagatctg ttctgcacaa 3600 agtggagtag tcagtcatcg atcaggaacc agacaccaga cttttattca tacagtgaag 3660 tgaagtgaag tgcagtgcag tgagttgctg gtttttgtac cacttagtat gtatttgtat 3720 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtgggtacc 3780 agcttgggct gagtggctcc ttcaacgttg cggttctgtc agttccaaac gtaaaacggc 3840 ttgtcccgcg tcatcggcgg gggtcataac gtgactccct taattctccg ctcatgatca 3900 gattgtcgtt tcccgccttc agtttaaact atcagtgttt gacaggatat attggcgggt 3960 aaacctaaga gaaaagagcg tttattagaa taacggatat ttaaaagggc gtgaaaaggt 4020 ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttcccctcg ggagtgcttg 4080 gcattccgta cgataatgac ttctgttcaa ccacccaaac gtcggaaagc ctgacgacgg 4140 agcagcattc caaaaagatc ccttggctcg tctgggtcgg ctagaaggtc gagtgggctg 4200 ctgtggcttg atccctcaac gcggtcgcgg acgtagcgca gcgccgaaaa atcctcgatc 4260 gcaaatccga cgctgtcgaa aagcgtgatc tgcttgtcgc tctttcggcc gacgtcctgg 4320 ccagtcatca cgcgccaaag ttccgtcaca ggatgatctg gcgcgagttg ctggatctcg 4380 ccttcaatcc gggtctgtgg cgggaactcc acgaaaatat ccgaacgcag caagatcgtc 4440 gaccaattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat 4500 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc 4560 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 4620 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc 4680 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt 4740 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct 4800 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac 4860 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact 4920 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa 4980 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga 5040 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt 5100 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga 5160 agccatacca aacgacgagc gtgacaccac gatgcctgca gggggggggg ggggggggac 5220 atgaggttgc cccgtattca gtgtcgctga tttgtattgt ctgaagttgt ttttacgtta 5280 agttgatgca gatcaattaa tacgatacct gcgtcataat tgattatttg acgtggtttg 5340 atggcctcca cgcacgttgt gatatgtaga tgataatcat tatcacttta cgggtccttt 5400 ccggtgatcc gacaggttac ggggcggcga cctcgcgggt tttcgctatt tatgaaaatt 5460 ttccggttta aggcgtttcc gttcttcttc gtcataactt aatgttttta tttaaaatac 5520 cctctgaaaa gaaaggaaac gacaggtgct gaaagcgagg ctttttggcc tctgtcgttt 5580 cctttctctg tttttgtccg tggaatgaac aatggaagtc cccccccccc cccccccctg 5640 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc 5700 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg 5760 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg 5820 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga 5880 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 5940 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa 6000 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 6060 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 6120 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 6180 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 6240 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc 6300 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 6360 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 6420 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 6480 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 6540 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 6600 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 6660 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 6720 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 6780 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 6840 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 6900 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca 6960 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct 7020 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg 7080 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat 7140 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatcc cccgatcaag 7200 tagatacact acatatatct acaatagaca tcgagccgga aggtgatgtt tactttcctg 7260 aaatccccag caattttagg ccagttttta cccaagactt cgcctctaac ataaattata 7320 gttaccaaat ctggcaaaag ggttaacaag tggcagcaac ggattcgcaa acctgtcacg 7380 ccttttgtgc caaaagccgc gccaggtttg cgatccgctg tgccaggcgt taggcgtcat 7440 atgaagattt cggtgatccc tgagcaggtg gcggaaacat tggatgctga gaaccatttc 7500 attgttcgtg aagtgttcga tgtgcaccta tccgaccaag gctttgaact atctaccaga 7560 agtgtgagcc cctaccggaa ggattacatc tcggatgatg actctgatga agactctgct 7620 tgctatggcg cattcatcga ccaagagctt gtcgggaaga ttgaactcaa ctcaacatgg 7680 aacgatctag cctctatcga acacattgtt gtgtcgcaca cgcaccgagg caaaggagtc 7740 gcgcacagtc tcatcgaatt tgcgaaaaag tgggcactaa gcagacagct ccttggcata 7800 cgattagaga cacaaacgaa caatgtacct gcctgcaatt tgtacgcaaa atgtggcttt 7860 actctcggcg gcattgacct gttcacgtat aaaactagac ctcaagtctc gaacgaaaca 7920 gcgatgtact ggtactggtt ctcgggagca caggatgacg cctaacaatt cattcaagcc 7980 gacaccgctt cgcggcgcgg cttaattcag gagttaaaca tcatgaggga agcggtgatc 8040 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg 8100 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt 8160 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg 8220 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta 8280 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa 8340 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg 8400 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta 8460 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta 8520 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta 8580 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat 8640 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa 8700 gctaggcagg cttatcttgg acaagaagat cgcttggcct cgcgcgcaga tcagttggaa 8760 gaatttgttc actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctaaca 8820 attcgttcaa gccgacgccg cttcgcggcg cggcttaact caagcgttag agagctgggg 8880 aagactatgc gcgatctgtt gaaggtggtt ctaagcctcg tacttgcgat ggcatcgggg 8940 caggcacttg ctgacctgcc aattgtttta gtggatgaag ctcgtcttcc ctatgactac 9000 tccccatcca actacgacat ttctccaagc aactacgaca actccataag caattacgac 9060 aatagtccat caaattacga caactctgag agcaactacg ataatagttc atccaattac 9120 gacaatagtc gcaacggaaa tcgtaggctt atatatagcg caaatgggtc tcgcactttc 9180 gccggctact acgtcattgc caacaatggg acaacgaact tcttttccac atctggcaaa 9240 aggatgttct acaccccaaa aggggggcgc ggcgtctatg gcggcaaaga tgggagcttc 9300 tgcggggcat tggtcgtcat aaatggccaa ttttcgcttg ccctgacaga taacggcctg 9360 aagatcatgt atctaagcaa ctagcctgct ctctaataaa atgttaggcc tcaacatcta 9420 gtcgcaagct gaggggaacc actagtgtca tacgaacctc caagagacgg ttacacaaac 9480 gggtacattg ttgatgtcat gtatgacaat cgcccaagta agtatccagc tgtgttcaga 9540 acgtacgtcc gaattaattc atcggggtac ggtcgacgat cgtcaacgtt cacttctaaa 9600 gaaatagcgc cactcagctt cctcagcggc tttatccagc gatttcctat tatgtcggca 9660 tagttctcaa gatcgacagc ctgtcacggt taagcgagaa atgaataaga aggctgataa 9720 ttcggatctc tgcgagggag atgatatttg atcacaggca gcaacgctct gtcatcgtta 9780 caatcaacat gctaccctcc gcgagatcat ccgtgtttca aacccggcag cttagttgcc 9840 gttcttccga atagcatcgg taacatgagc aaagtctgcc gccttacaac ggctctcccg 9900 ctgacgccgt cccggactga tgggctgcct gtatcgagtg gtgattttgt gccgagctgc 9960 cggtcgggga gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg 10020 cttagacaac ttaataacac attgcggacg tttttaatgt actgaattgt ctagacccgg 10080 ggatctcatg tttgacagct tatcatcgga tctagtaaca tagatgacac cgcgcgcgat 10140 aatttatcct agtttgcgcg ctatattttg ttttctatcg cgtattaaat gtataattgc 10200 gggactctaa tcataaaaac ccatctcata aataacgtca tgcattacat gttaattatt 10260 acatgcttaa cgtaattcaa cagaaattag atgataatca tcgcaagacc ggcaacagga 10320 ttcaatctta agaaacttta ttgccaaatg tttgaacgat ctctgcaggt cgacggatcg 10380 agctcccagc ttagcaagag atgttaattt tttcagtaag ctcttacagc ttgttgtaaa 10440 cacgcgctaa acggccgtgg cctttgacag tcaccggtga ttcgttggcg gcaataaacg 10500 ctgattcacc cggtttaagc tgtaactgct gagaaccttt ccacaacgtt gcatcgcctt 10560 cgacgcagaa caaaatggcg gcactctgct ggctaatggt ggtttcttta tcactaaggt 10620 catgcagcga gaaggcaaaa tcatccactg gaatcgggaa gtccagttct gcaccttgtt 10680 tcaccggctg ggtcaacaac tggttagccg gtttggcttc gaatttcaca ttggcaacca 10740 gttccggaat atcaatgtat ttaggcgtca gacccgcacg cagcacgtta tcggagtttg 10800 ccatcacttc cagcgccacg ccttgcaggt aagcgtgcgg tgtttcagcg aacaggaaca 10860 tcgcttcgcc agggttcaat ttcaccacat tcagcaatag cggggagaac agaccgctgt 10920 cttccgggta aaattcagaa attaaacgaa tcgtttgcca cggttcaccc tgctggctat 10980 cgagggccga ttttaaaatc gccagcgcgc gggatttttc ttcaccctgc atattcaaca 11040 ggctggcgaa cagttcgctt aaacgttcgg catcaggctg ttgtaaaaag tgagcaatcg 11100 ccggatgtgc acctgcgacc ggctggagta gggagacaat ctcggaaaat tcacgaaacg 11160 cgttcatcgc aaggaaaggc gtcagcgcaa aaaccagctc cggcttgtgg ttaggatctt 11220 tatagttacg ctcggcggca tccatcggga tacctgcggc attttctttg gcaaaaccga 11280 tttcagaatt gtgtttgttt ggatgaacct gaatggagag tggctgtgct gcgcataata 11340 ctttgaacag gaaaggcagt tcgccaaagc gtttggcaac ggcctctccg agcagagtcg 11400 atttatcact ctcaatcaca tcacgcagtg aaacgatatc tccggcggca ttctgcactc 11460 gtgaactgct tttcggatgt gcgcccatcc acagctcggc catcggctgg ctggacggat 11520 tttccatacc ataaagttca gtcaacgcgt tttgctgccc caggcatagt tttgcactga 11580 gttaatgagt ttttgcatga tcggggatcc ctgcagaagt aacaccaaac aacagggtga 11640 gcatcgacaa aagaaacagt accaagcaaa taaatagcgt atgaaggcag ggctaaaaaa 11700 atccacatat agctgctgca tatgccatca tccaagtata tcaagatcaa aataattata 11760 aaacatactt gtttattata atagataggt actcaaggtt agagcatatg aatagatgct 11820 gcatatgcca tcatgtatat gcatcagtaa aacccacatc aacatgtata cctatcctag 11880 atcgatattt ccatccatct taaactcgta actatgaaga tgtatgacac acacatacag 11940 ttccaaaatt aataaataca ccaggtagtt tgaaacggcg tctactccga tctagaacga 12000 atgaacgacc gcccaaccac accacatcat cacaaccaag cgaacaaaaa gcatctctgt 12060 atatgcatca gtaaaacccg catcaacatg tatacctatc ctagatcgat atttccatcc 12120 atcatcttca attcgtaact atgaatatgt atggcacaca catacagatc caaaattaat 12180 aaatccacca ggtagtttga aacagaattc tactccgatc tagaacgacc gcccaaccag 12240 accacatcat cacaaccaag acaaaaaaaa gcatgaaaag atgacccgac aaacaagtgc 12300 acggcatata ttgaaataaa ggaaaagggc aaaccaaacc ctatgcaacg aaacaaaaaa 12360 aatcatgaaa tcgatcccgt ctgcggaacg gctagagcca tcccaggatt ccccaaagag 12420 aaacactggc aagttagcaa tcagaacgtg tctgacgtac aggtcgcatc cgtgtacgaa 12480 cgctagcagc acggatctaa cacaaacacg gatctaacac aaacatgaac agaagtagaa 12540 ctaccgggcc ctaaccatgg accggaacgc cgatctagag aaggtagaga gggggggggg 12600 gggaggacga gcggcgtacc ttgaagcgga ggtgccgacg ggtggatttg ggggagatct 12660 ggttgtgtgt gtgtgcgctc cgaacaacac gaggttgggg aaagagggtg tggagggggt 12720 gtctatttat tacggcgggc gaggaaggga aagcgaagga gcggtgggaa aggaatcccc 12780 cgtagctgcc gtgccgtgag aggaggagga ggccgcctgc cgtgccggct cacgtctgcc 12840 gctccgccac gcaatttctg gatgccgaca gcggagcaag tccaacggtg gagcggaact 12900 ctcgagaggg gtccagaggc agcgacagag atgccgtgcc gtctgcttcg cttggcccga 12960 cgcgacgctg ctggttcgct ggttggtgtc cgttagactc gtcgacggcg tttaacaggc 13020 tggcattatc tactcgaaac aagaaaaatg tttccttagt ttttttaatt tcttaaaggg 13080 tatttgttta atttttagtc actttatttt attctatttt atatctaaat tattaaataa 13140 aaaaactaaa atagagtttt agttttctta atttagaggc taaaatagaa taaaatagat 13200 gtactaaaaa aattagtcta taaaaaccat taaccctaaa ccctaaatgg atgtactaat 13260 aaaatggatg aagtattata taggtgaagc tatttgcaaa aaaaaaggag aacacatgca 13320 cactaaaaag ataaaactgt agagtcctgt tgtcaaaata ctcaattgtc ctttagacca 13380 tgtctaactg ttcatttata tgattctcta aaacactgat attattgtag tactatagat 13440 tatattattc gtagagtaaa gtttaaatat atgtataaag atagataaac tgcacttcaa 13500 acaagtgtga caaaaaaaat atgtggtaat tttttataac ttagacatgc aatgctcatt 13560 atctctagag aggggcacga ccgggtcacg ctgcactgca ggcatgcaag cttgcacatg 13620 acaacaattg taagaggatg gagaccacaa cgatccaaca atacttctgc gacgggctgt 13680 gaagtataga gaagttaaac gcccaaaagc cattgtgttt ggaattttta gttattctat 13740 ttttcatgat gtatcttcct ctaacatgcc ttaatttgca aatttggtat aactactgat 13800 tgaaaatata tgtatgtaaa aaaatactaa gcatatttgt gaagctaaac atgatgttat 13860 ttaagaaaat atgttgttaa cagaataaga ttaatatcga aatggaaaca tctgtaaatt 13920 agaatcatct tacaagctaa gagatgttca cgctttgaga aacttcttca gatcatgacc 13980 gtagaagtag ctctccaaga ctcaacgaag gctgctgcaa ttccacaaat gcatgacatg 14040 catccttgta accgtcgtcg ccgctataaa cacggataac tcaattccct gctccatcaa 14100 tttagaaatg agcaagcaag cacccgatcg ctcaccccat atgcaccaat ctgactccca 14160 agtctctgtt tcgcattagt accgccagca ctccacctat agctaccaat tgagaccttt 14220 ccagcctaag cagatcgatt gatcgttaga gtcaaagagt tggtggtacg ggtactttaa 14280 ctaccatgga atgatggggc gtgatgtaga gcggaaagcg cctccctacg cggaacaaca 14340 ccctcgccat gccgctcgac tacagcctcc tcctcgtcgg ccgcccacaa cgagggagcc 14400 cgtggtcgca gccaccgacc agcatgtctc tgtgtcctcg tccgacctcg acatgtcatg 14460 gcaaacagtc ggacgccagc accagactga cgacatgagt ctctgaagag cccgccacct 14520 agaaagatcc gagccctgct gctggtagtg gtaaccattt tcgtcgcgct gacgcggaga 14580 gcgagaggcc agaaatttat agcgactgac gctgtggcag gcacgctatc ggaggttacg 14640 acgtggcggg tcactcgacg cggagttcac aggtcctatc cttgcatcgc tcgggccgga 14700 gtttacggga cttatcctta cgacgtgctc taaggttgcg ataacgggcg gaggaaggcg 14760 tgtggcgtgc ggagacggtt tatacacgta gtgtgcggga gtgtgtttcg tagacgcggg 14820 aaagcacgac gacttacgaa ggttagtgga ggaggaggac acactaaaat caggacgcaa 14880 gaaactcttc tattatagta gtagagaaga gattatagga gtgtgggttg attctaaaga 14940 aaatcgacgc aggacaaccg tcaaaacggg tgctttaata tagtagatat atatatatag 15000 agagagagag aaagtacaaa ggatgcattt gtgtctgcat atgatcggag tattactaac 15060 ggccgtcgta agaaggtcca tcatgcgtgg agcgagccca tttggttggt tgtcaggccg 15120 cagttaaggc ctccatatat gattgtcgtc gggcccataa cagcatctcc tccaccagtt 15180 tattgtaaga ataaattaag tagagatatt tgtcgtcggg cagaagaaac ttggacaaga 15240 agaagaagca agctaggcca atttcttgcc ggcaagagga agatagtggc ctctagttta 15300 tatatcggcg tgatgatgat gctcctagct agaaatgaga gaagaaaaac ggacgcgtgt 15360 ttggtgtgtg tcaatggcgt ccatccttcc atcagatcag aacgatgaaa aagtcaagca 15420 cggcatgcat agtatatgta tagcttgttt tagtgtggct ttgctgagac gaatgaaagc 15480 aacggcgggc atatttttca gtggctgtag ctttcaggct gaaagagacg tggcatgcaa 15540 taattcaggg aattcgtcag ccaattgagg tagctagtca acttgtacat tggtgcgagc 15600 aattttccgc actcaggagg gctagtttga gagtccaaaa actataggag attaaagagg 15660 ctaaaatcct ctccttattt aattttaaat aagtagtgta tttgtatttt aactcctcca 15720 acccttccga ttttatggct ctcaaactag cattcagtct aatgcatgca tgcttggcta 15780 gaggtcgtat ggggttgtta atagcatagc tagctacaag ttaaccgggt cttttatatt 15840 taataaggac aggcaaagta ttacttacaa ataaagaata aagctaggac gaactcgtgg 15900 attattacta aatcgaaatg gacgtaatat tccaggcaag aataattgtt cgatcaggag 15960 acaagtgggg cattggaccg gttcttgcaa gcaagagcct atggcgtggt gacacggcgc 16020 gttgcccata catcatgcct ccatcgatga tccatcctca cttgctataa aaagaggtgt 16080 ccatggtgct caagctcagc caagcaaata agacgacttg tttcattgat tcttcaagag 16140 atcgagcttc ttttgcacca caaggtcgag gatccaaca 16179 <210> 13 <211> 15643 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: pNOV1441 <220> <221> misc_feature (222) (14) .. (1414) <223> Maize ubiquitin (Mz Ubi) promoter <220> <221> misc_feature (222) (2037) .. (5618) <223> synthetic nucleotide sequence encoding the toxin       portion of H04 plus a full-length Cry1Ab tail       portion <220> <221> misc_feature (222) (5821) .. (6711) <223> Mz Ubi promoter <220> <221> misc_feature (222) (7831) .. (9039) <223> PMI <400> 13 aagctggtac aagcttgcat gcctgcagtg cagcgtgacc cggtcgtgcc cctctctaga 60 gataatgagc attgcatgtc taagttataa aaaattacca catatttttt ttgtcacact 120 tgtttgaagt gcagtttatc tatctttata catatattta aactttactc tacgaataat 180 ataatctata gtactacaat aatatcagtg ttttagagaa tcatataaat gaacagttag 240 acatggtcta aaggacaatt gagtattttg acaacaggac tctacagttt tatcttttta 300 gtgtgcatgt gttctccttt ttttttgcaa atagcttcac ctatataata cttcatccat 360 tttattagta catccattta gggtttaggg ttaatggttt ttatagacta atttttttag 420 tacatctatt ttattctatt ttagcctcta aattaagaaa actaaaactc tattttagtt 480 tttttattta ataatttaga tataaaatag aataaaataa agtgactaaa aattaaacaa 540 atacccttta agaaattaaa aaaactaagg aaacattttt cttgtttcga gtagataatg 600 ccagcctgtt aaacgccgtc gacgagtcta acggacacca accagcgaac cagcagcgtc 660 gcgtcgggcc aagcgaagca gacggcacgg catctctgtc gctgcctctg gacccctctc 720 gagagttccg ctccaccgtt ggacttgctc cgctgtcggc atccagaaat tgcgtggcgg 780 agcggcagac gtgagccggc acggcaggcg gcctcctcct cctctcacgg cacggcagct 840 acgggggatt cctttcccac cgctccttcg ctttcccttc ctcgcccgcc gtaataaata 900 gacaccccct ccacaccctc tttccccaac ctcgtgttgt tcggagcgca cacacacaca 960 accagatctc ccccaaatcc acccgtcggc acctccgctt caaggtacgc cgctcgtcct 1020 cccccccccc ccctctctac cttctctaga tcggcgttcc ggtccatggt tagggcccgg 1080 tagttctact tctgttcatg tttgtgttag atccgtgttt gtgttagatc cgtgctgcta 1140 gcgttcgtac acggatgcga cctgtacgtc agacacgttc tgattgctaa cttgccagtg 1200 tttctctttg gggaatcctg ggatggctct agccgttccg cagacgggat cgatttcatg 1260 attttttttg tttcgttgca tagggtttgg tttgcccttt tcctttattt caatatatgc 1320 cgtgcacttg tttgtcgggt catcttttca tgcttttttt tgtcttggtt gtgatgatgt 1380 ggtctggttg ggcggtcgtt ctagatcgga gtagaattct gtttcaaact acctggtgga 1440 tttattaatt ttggatctgt atgtgtgtgc catacatatt catagttacg aattgaagat 1500 gatggatgga aatatcgatc taggataggt atacatgttg atgcgggttt tactgatgca 1560 tatacagaga tgctttttgt tcgcttggtt gtgatgatgt ggtgtggttg ggcggtcgtt 1620 cattcgttct agatcggagt agaatactgt ttcaaactac ctggtgtatt tattaatttt 1680 ggaactgtat gtgtgtgtca tacatcttca tagttacgag tttaagatgg atggaaatat 1740 cgatctagga taggtataca tgttgatgtg ggttttactg atgcatatac atgatggcat 1800 atgcagcatc tattcatatg ctctaacctt gagtacctat ctattataat aaacaagtat 1860 gttttataat tattttgatc ttgatatact tggatgatgg catatgcagc agctatatgt 1920 ggattttttt agccctgcct tcatacgcta tttatttgct tggtactgtt tcttttgtcg 1980 atgctcaccc tgttgtttgg tgttacttct gcaggtcgac tctagaggat ccaacaatgg 2040 acaacaaccc caacatcaac gagtgcatcc cctacaactg cctgagcaac cccgaggtgg 2100 aggtgctggg cggcgagcgc atcgagaccg gctacacccc catcgacatc agcctgagcc 2160 tgacccagtt cctgctgagc gagttcgtgc ccggcgccgg cttcgtgctg ggcctggtgg 2220 acatcatctg gggcatcttc ggccccagcc agtgggacgc cttcctggtg cagatcgagc 2280 agttgataaa ccaacgcata gaggaattcg cccgcaacca ggccatcagc cgcctggagg 2340 gcctgagcaa cctgtaccaa atctacgccg agagcttccg cgagtgggag gccgacccca 2400 ccaaccccgc cctgcgcgag gagatgcgca tccagttcaa cgacatgaac agcgccctga 2460 ccaccgccat ccccctgttc gccgtgcaga actaccaggt gcccctgctg agcgtgtacg 2520 tgcaggccgc caacctgcac ctgagcgtgc tgcgcgacgt cagcgtgttc ggccagcgct 2580 ggggcttcga cgccgccacc atcaacagcc gctacaacga cctgacccgc ctgatcggca 2640 actacaccga ccacgccgtg cgctggtaca acaccggcct ggagcgcgtg tggggtcccg 2700 acagccgcga ctggatcagg tacaaccagt tccgccgcga gctgaccctg accgtgctgg 2760 acatcgtgag cctgttcccc aactacgaca gccgcaccta ccccatccgc accgtgagcc 2820 agctgacccg cgagatttac accaaccccg tgctggagaa cttcgacggc agcttccgcg 2880 gcagcgccca gggcatcgag ggcagcatcc gcagccccca cctgatggac atcctgaaca 2940 gcatcaccat ctacaccgac gcccaccgcg gcgagtacta ctggagcggc caccagatca 3000 tggccagccc cgtcggcttc agcggccccg agttcacctt ccccctgtac ggcaccatgg 3060 gcaacgctgc acctcagcag cgcatcgtgg cacagctggg ccagggagtg taccgcaccc 3120 tgagcagcac cctgtaccgt cgacctttca acatcggcat caacaaccag cagctgagcg 3180 tgctggacgg caccgagttc gcctacggca ccagcagcaa cctgcccagc gccgtgtacc 3240 gcaagagcgg caccgtggac agcctggacg agatcccccc tcagaacaac aacgtgccac 3300 ctcgacaggg cttcagccac cgtctgagcc acgtgagcat gttccgcagt ggcttcagca 3360 acagcagcgt gagcatcatc cgtgcaccca tgttcagctg gattcaccgc agcgccaccc 3420 tgaccaacac catcgacccc gagcgcatca accagatccc cctggtgaag ggcttccggg 3480 tgtggggcgg caccagcgtg atcaccggcc ccggcttcac cggaggcgac atcctgcgca 3540 gaaacacctt cggcgacttc gtgagcctgc aggtgaacat caacagcccc atcacccagc 3600 gttaccgcct gcgcttccgc tacgccagca gccgcgacgc ccgtgtgatc gtgctgactg 3660 gcgccgctag caccggtgtg ggcggtcagg tgagcgtgaa catgcccctg cagaagacta 3720 tggagatcgg cgagaacctg actagtcgca ccttccgcta caccgacttc agcaacccct 3780 tcagcttccg cgccaacccc gacatcatcg gcatcagcga gcagcccctg ttcggtgccg 3840 gcagcatcag cagcggcgag ctgtacatcg acaagatcga gatcatcctg gccgacgcca 3900 ccttcgaggc cgagagcgac ctggagcgcg cccagaaggc cgtgaacgcc ctgttcacca 3960 gcagcaacca gatcggcctg aagaccgacg tgaccgacta ccacatcgac caggtgagca 4020 acctggtgga ctgcttaagc gacgagttct gcctggacga gaagaaggag ctgagcgaga 4080 aggtgaagca cgccaagcgc ctgagcgacg agcgcaacct gctgcaggac cccaacttcc 4140 gcggcatcaa ccgccagctg gaccgcggct ggcgaggcag caccgatatc accatccagg 4200 gcggcgacga cgtgttcaag gagaactacg tgaccctgca gggcaccttc gacgagtgct 4260 accccaccta cctgtaccag ccgatcgacg agagcaagct gaaggcctac acccgctacc 4320 agctgcgcgg ctacatcgag gacagccagg acctggaaat ctacctgatc cgctacaacg 4380 cgaagcacga gaccgtgaac gtgcccggca ccggcagcct gtggcccccg agcgccccca 4440 gccccatcgg caagtgcggg gagccgaatc gatgcgctcc gcacctggag tggaacccgg 4500 acctagactg cagctgcagg gacggggaga agtgcgccca ccacagccac cacttcagcc 4560 tggacatcga cgtgggctgc accgacctga acgaggacct gggcgtgtgg gtgatcttca 4620 agatcaagac ccaggacggc cacgcccgcc tgggcaatct agagttcctg gaggagaagc 4680 ccctggtggg cgaggccctg gcccgcgtga agcgtgctga gaagaagtgg cgcgacaagc 4740 gcgagaagct ggagtgggag accaacatcg tgtacaagga ggccaaggag agcgtggacg 4800 ccctgttcgt gaacagccag tacgaccgcc tgcaggccga caccaacatc gccatgatcc 4860 acgccgccga caagcgcgtg cacagcattc gcgaggccta cctgcccgag ctgagcgtga 4920 tccccggtgt gaacgccgcc atcttcgagg aactcgaggg ccgcatcttc accgccttca 4980 gcctgtacga cgcccgcaac gtgatcaaga acggcgactt caacaacggc ctgagctgct 5040 ggaacgtgaa gggccacgtg gacgtggagg agcagaacaa ccaccgcagc gtgctggtgg 5100 tgcccgagtg ggaggccgag gtgagccagg aggtgcgcgt gtgccccggc cgcggctaca 5160 tcctgcgcgt gaccgcctac aaggagggct acggcgaggg ctgcgtgacc atccacgaga 5220 tcgagaacaa caccgacgag ctcaagttca gcaactgcgt ggaggaggag gtttacccca 5280 acaacaccgt gacctgcaac gactacaccg cgacccagga ggagtacgaa ggcacctaca 5340 cctctcgcaa caggggttac gacggcgcct acgagtccaa cagctccgtg ccagctgact 5400 acgccagcgc ccacgaggag aaagcctaca ccgacggtag acgcgacaac ccatgtgaga 5460 gcaacagagg ctacggcgac tacacccccc tgcccgctgg atacgtgacc aaggagctgg 5520 agtacttccc cgagaccgac aaggtgtgga tcgagattgg cgagaccgag ggcaccttca 5580 tcgtggacag cgtggagctg ctgctgatgg aggagtagta gatctgttct gcacaaagtg 5640 gagtagtcag tcatcgatca ggaaccagac accagacttt tattcataca gtgaagtgaa 5700 gtgaagtgca gtgcagtgag ttgctggttt ttgtaccact tagtatgtat ttgtatttgt 5760 aaaatacttc tatcaataaa atttctaatt cctaaaacca aaatccagtg ggtaccagct 5820 tgcatgcctg cagtgcagcg tgacccggtc gtgcccctct ctagagataa tgagcattgc 5880 atgtctaagt tataaaaaat taccacatat tttttttgtc acacttgttt gaagtgcagt 5940 ttatctatct ttatacatat atttaaactt tactctacga ataatataat ctatagtact 6000 acaataatat cagtgtttta gagaatcata taaatgaaca gttagacatg gtctaaagga 6060 caattgagta ttttgacaac aggactctac agttttatct ttttagtgtg catgtgttct 6120 cctttttttt tgcaaatagc ttcacctata taatacttca tccattttat tagtacatcc 6180 atttagggtt tagggttaat ggtttttata gactaatttt tttagtacat ctattttatt 6240 ctattttagc ctctaaatta agaaaactaa aactctattt tagttttttt atttaataat 6300 ttagatataa aatagaataa aataaagtga ctaaaaatta aacaaatacc ctttaagaaa 6360 ttaaaaaaac taaggaaaca tttttcttgt ttcgagtaga taatgccagc ctgttaaacg 6420 ccgtcgacga gtctaacgga caccaaccag cgaaccagca gcgtcgcgtc gggccaagcg 6480 aagcagacgg cacggcatct ctgtcgctgc ctctggaccc ctctcgagag ttccgctcca 6540 ccgttggact tgctccgctg tcggcatcca gaaattgcgt ggcggagcgg cagacgtgag 6600 ccggcacggc aggcggcctc ctcctcctct cacggcacgg cagctacggg ggattccttt 6660 cccaccgctc cttcgctttc ccttcctcgc ccgccgtaat aaatagacac cccctccaca 6720 ccctctttcc ccaacctcgt gttgttcgga gcgcacacac acacaaccag atctccccca 6780 aatccacccg tcggcacctc cgcttcaagg tacgccgctc gtcctccccc cccccccctc 6840 tctaccttct ctagatcggc gttccggtcc atggttaggg cccggtagtt ctacttctgt 6900 tcatgtttgt gttagatccg tgtttgtgtt agatccgtgc tgctagcgtt cgtacacgga 6960 tgcgacctgt acgtcagaca cgttctgatt gctaacttgc cagtgtttct ctttggggaa 7020 tcctgggatg gctctagccg ttccgcagac gggatcgatt tcatgatttt ttttgtttcg 7080 ttgcataggg tttggtttgc ccttttcctt tatttcaata tatgccgtgc acttgtttgt 7140 cgggtcatct tttcatgctt ttttttgtct tggttgtgat gatgtggtct ggttgggcgg 7200 tcgttctaga tcggagtaga attctgtttc aaactacctg gtggatttat taattttgga 7260 tctgtatgtg tgtgccatac atattcatag ttacgaattg aagatgatgg atggaaatat 7320 cgatctagga taggtataca tgttgatgcg ggttttactg atgcatatac agagatgctt 7380 tttgttcgct tggttgtgat gatgtggtgt ggttgggcgg tcgttcattc gttctagatc 7440 ggagtagacg ccgtttcaaa ctacctggtg tatttattaa ttttggaact gtatgtgtgt 7500 gtcatacatc ttcatagtta cgagtttaag atggatggaa atatcgatct aggataggta 7560 tacatgttga tgtgggtttt actgatgcat atacatgatg gcatatgcag catctattca 7620 tatgctctaa ccttgagtac ctatctatta taataaacaa gtatgtttta taattatttt 7680 gatcttgata tacttggatg atggcatatg cagcagctat atgtggattt ttttagccct 7740 gccttcatac gctatttatt tgcttggtac tgtttctttt gtcgatgctc accctgttgt 7800 ttggtgttac ttctgcaggg atccccgatc atgcaaaaac tcattaactc agtgcaaaac 7860 tatgcctggg gcagcaaaac gcgttgactg aactttatgg tatggaaaat ccgtccagcc 7920 agccgatggc cgagctgtgg atgggcgcac atccgaaaag cagttcacga gtgcagaatg 7980 ccgccggaga tatcgtttca ctgcgtgatg tgattgagag tgataaatcg actctgctcg 8040 gagaggccgt tgccaaacgc tttggcgaac tgcctttcct gttcaaagta ttatgcgcag 8100 cacagccact ctccattcag gttcatccaa acaaacacaa ttctgaaatc ggttttgcca 8160 aagaaaatgc cgcaggtatc ccgatggatg ccgccgagcg taactataaa gatcctaacc 8220 acaagccgga gctggttttt gcgctgacgc ctttccttgc gatgaacgcg tttcgtgaat 8280 tttccgagat tgtctcccta ctccagccgg tcgcaggtgc acatccggcg attgctcact 8340 ttttacaaca gcctgatgcc gaacgtttaa gcgaactgtt cgccagcctg ttgaatatgc 8400 agggtgaaga aaaatcccgc gcgctggcga ttttaaaatc ggccctcgat agccagcagg 8460 gtgaaccgtg gcaaacgatt cgtttaattt ctgaatttta cccggaagac agcggtctgt 8520 tctccccgct attgctgaat gtggtgaaat tgaaccctgg cgaagcgatg ttcctgttcg 8580 ctgaaacacc gcacgcttac ctgcaaggcg tggcgctgga agtgatggca aactccgata 8640 acgtgctgcg tgcgggtctg acgcctaaat acattgatat tccggaactg gttgccaatg 8700 tgaaattcga agccaaaccg gctaaccagt tgttgaccca gccggtgaaa caaggtgcag 8760 aactggactt cccgattcca gtggatgatt ttgccttctc gctgcatgac cttagtgata 8820 aagaaaccac cattagccag cagagtgccg ccattttgtt ctgcgtcgaa ggcgatgcaa 8880 cgttgtggaa aggttctcag cagttacagc ttaaaccggg tgaatcagcg tttattgccg 8940 ccaacgaatc accggtgact gtcaaaggcc acggccgttt agcgcgtgtt tacaacaagc 9000 tgtaagagct tactgaaaaa attaacatct cttgctaagc tgggagctcg atccgtcgac 9060 ctgcagagat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg 9120 tcttgcgatg attatcatct aatttctgtt gaattacgtt aagcatgtaa taattaacat 9180 gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat 9240 ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt 9300 gtcatctatg ttactagatc cgatgataag ctgtcaaaca tgagatcccc gggtctagac 9360 aattcagtac attaaaaacg tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt 9420 tacaccacaa tatatcctgc caccagccag ccaacagctc cccgaccggc agctcggcac 9480 aaaatcacca ctcgatacag gcagcccatc agtccgggac ggcgtcagcg ggagagccgt 9540 tgtaaggcgg cagactttgc tcatgttacc gatgctattc ggaagaacgg caactaagct 9600 gccgggtttg aaacacggat gatctcgcgg agggtagcat gttgattgta acgatgacag 9660 agcgttgctg cctgtgatca aatatcatct ccctcgcaga gatccgaatt atcagccttc 9720 ttattcattt ctcgcttaac cgtgacaggc tgtcgatctt gagaactatg ccgacataat 9780 aggaaatcgc tggataaagc cgctgaggaa gctgagtggc gctatttctt tagaagtgaa 9840 cgttgacgat cgtcgaccgt accccgatga attaattcgg acgtacgttc tgaacacagc 9900 tggatactta cttgggcgat tgtcatacat gacatcaaca atgtacccgt ttgtgtaacc 9960 gtctcttgga ggttcgtatg acactagtgg ttcccctcag cttgcgacta gatgttgagg 10020 cctaacattt tattagagag caggctagtt gcttagatac atgatcttca ggccgttatc 10080 tgtcagggca agcgaaaatt ggccatttat gacgaccaat gccccgcaga agctcccatc 10140 tttgccgcca tagacgccgc gccccccttt tggggtgtag aacatccttt tgccagatgt 10200 ggaaaagaag ttcgttgtcc cattgttggc aatgacgtag tagccggcga aagtgcgaga 10260 cccatttgcg ctatatataa gcctacgatt tccgttgcga ctattgtcgt aattggatga 10320 actattatcg tagttgctct cagagttgtc gtaatttgat ggactattgt cgtaattgct 10380 tatggagttg tcgtagttgc ttggagaaat gtcgtagttg gatggggagt agtcataggg 10440 aagacgagct tcatccacta aaacaattgg caggtcagca agtgcctgcc ccgatgccat 10500 cgcaagtacg aggcttagaa ccaccttcaa cagatcgcgc atagtcttcc ccagctctct 10560 aacgcttgag ttaagccgcg ccgcgaagcg gcgtcggctt gaacgaattg ttagacatta 10620 tttgccgact accttggtga tctcgccttt cacgtagtga acaaattctt ccaactgatc 10680 tgcgcgcgag gccaagcgat cttcttgtcc aagataagcc tgcctagctt caagtatgac 10740 gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc 10800 gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc 10860 atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa 10920 tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac 10980 gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc 11040 gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc 11100 tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag 11160 aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg 11220 cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt 11280 caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg 11340 gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc 11400 cctcatgatg tttaactcct gaattaagcc gcgccgcgaa gcggtgtcgg cttgaatgaa 11460 ttgttaggcg tcatcctgtg ctcccgagaa ccagtaccag tacatcgctg tttcgttcga 11520 gacttgaggt ctagttttat acgtgaacag gtcaatgccg ccgagagtaa agccacattt 11580 tgcgtacaaa ttgcaggcag gtacattgtt cgtttgtgtc tctaatcgta tgccaaggag 11640 ctgtctgctt agtgcccact ttttcgcaaa ttcgatgaga ctgtgcgcga ctcctttgcc 11700 tcggtgcgtg tgcgacacaa caatgtgttc gatagaggct agatcgttcc atgttgagtt 11760 gagttcaatc ttcccgacaa gctcttggtc gatgaatgcg ccatagcaag cagagtcttc 11820 atcagagtca tcatccgaga tgtaatcctt ccggtagggg ctcacacttc tggtagatag 11880 ttcaaagcct tggtcggata ggtgcacatc gaacacttca cgaacaatga aatggttctc 11940 agcatccaat gtttccgcca cctgctcagg gatcaccgaa atcttcatat gacgcctaac 12000 gcctggcaca gcggatcgca aacctggcgc ggcttttggc acaaaaggcg tgacaggttt 12060 gcgaatccgt tgctgccact tgttaaccct tttgccagat ttggtaacta taatttatgt 12 120 tagaggcgaa gtcttgggta aaaactggcc taaaattgct ggggatttca ggaaagtaaa 12180 catcaccttc cggctcgatg tctattgtag atatatgtag tgtatctact tgatcggggg 12240 atctgctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 12300 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 12360 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga 12420 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc 12480 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt 12540 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 12600 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 12660 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 12720 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 12780 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 12840 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 12900 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 12960 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 13020 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 13080 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 13140 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 13200 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 13260 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 13320 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 13380 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 13440 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 13500 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 13560 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 13620 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 13680 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 13740 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca gggggggggg 13800 ggggggggga cttccattgt tcattccacg gacaaaaaca gagaaaggaa acgacagagg 13860 ccaaaaagcc tcgctttcag cacctgtcgt ttcctttctt ttcagagggt attttaaata 13920 aaaacattaa gttatgacga agaagaacgg aaacgcctta aaccggaaaa ttttcataaa 13980 tagcgaaaac ccgcgaggtc gccgccccgt aacctgtcgg atcaccggaa aggacccgta 14040 aagtgataat gattatcatc tacatatcac aacgtgcgtg gaggccatca aaccacgtca 14100 aataatcaat tatgacgcag gtatcgtatt aattgatctg catcaactta acgtaaaaac 14160 aacttcagac aatacaaatc agcgacactg aatacggggc aacctcatgt cccccccccc 14220 cccccccctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 14280 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 14340 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 14400 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 14460 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 14520 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 14580 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 14640 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 14700 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 14760 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 14820 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 14880 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc 14940 tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattggtcga cgatcttgct 15000 gcgttcggat attttcgtgg agttcccgcc acagacccgg attgaaggcg agatccagca 15060 actcgcgcca gatcatcctg tgacggaact ttggcgcgtg atgactggcc aggacgtcgg 15120 ccgaaagagc gacaagcaga tcacgctttt cgacagcgtc ggatttgcga tcgaggattt 15180 ttcggcgctg cgctacgtcc gcgaccgcgt tgagggatca agccacagca gcccactcga 15240 ccttctagcc gacccagacg agccaaggga tctttttgga atgctgctcc gtcgtcaggc 15300 tttccgacgt ttgggtggtt gaacagaagt cattatcgta cggaatgcca agcactcccg 15360 aggggaaccc tgtggttggc atgcacatac aaatggacga acggataaac cttttcacgc 15420 ccttttaaat atccgttatt ctaataaacg ctcttttctc ttaggtttac ccgccaatat 15480 atcctgtcaa acactgatag tttaaactga aggcgggaaa cgacaatctg atcatgagcg 15540 gagaattaag ggagtcacgt tatgaccccc gccgatgacg cgggacaagc cgttttacgt 15600 ttggaactga cagaaccgca acgttgaagg agccactcag ccc 15643 <210> 14 <211> 15503 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: pNOV1305 <220> <221> misc_feature (222) (1) .. (3582) <223> synthetic nucleotide sequence encoding the toxin       portion of H04 plus a full-length Cry1Ab tail       portion <220> <221> misc_feature (222) (3790) .. (5771) <223> Zm Ubi promoter <220> <221> misc_feature (222) (5868) .. (6971) <223> PMI <220> <221> misc_feature (222) (12934) .. (15494) <223> MTL promoter <400> 14 atggacaaca accccaacat caacgagtgc atcccctaca actgcctgag caaccccgag 60 gtggaggtgc tgggcggcga gcgcatcgag accggctaca cccccatcga catcagcctg 120 agcctgaccc agttcctgct gagcgagttc gtgcccggcg ccggcttcgt gctgggcctg 180 gtggacatca tctggggcat cttcggcccc agccagtggg acgccttcct ggtgcagatc 240 gagcagttga taaaccaacg catagaggaa ttcgcccgca accaggccat cagccgcctg 300 gagggcctga gcaacctgta ccaaatctac gccgagagct tccgcgagtg ggaggccgac 360 cccaccaacc ccgccctgcg cgaggagatg cgcatccagt tcaacgacat gaacagcgcc 420 ctgaccaccg ccatccccct gttcgccgtg cagaactacc aggtgcccct gctgagcgtg 480 tacgtgcagg ccgccaacct gcacctgagc gtgctgcgcg acgtcagcgt gttcggccag 540 cgctggggct tcgacgccgc caccatcaac agccgctaca acgacctgac ccgcctgatc 600 ggcaactaca ccgaccacgc cgtgcgctgg tacaacaccg gcctggagcg cgtgtggggt 660 cccgacagcc gcgactggat caggtacaac cagttccgcc gcgagctgac cctgaccgtg 720 ctggacatcg tgagcctgtt ccccaactac gacagccgca cctaccccat ccgcaccgtg 780 agccagctga cccgcgagat ttacaccaac cccgtgctgg agaacttcga cggcagcttc 840 cgcggcagcg cccagggcat cgagggcagc atccgcagcc cccacctgat ggacatcctg 900 aacagcatca ccatctacac cgacgcccac cgcggcgagt actactggag cggccaccag 960 atcatggcca gccccgtcgg cttcagcggc cccgagttca ccttccccct gtacggcacc 1020 atgggcaacg ctgcacctca gcagcgcatc gtggcacagc tgggccaggg agtgtaccgc 1080 accctgagca gcaccctgta ccgtcgacct ttcaacatcg gcatcaacaa ccagcagctg 1140 agcgtgctgg acggcaccga gttcgcctac ggcaccagca gcaacctgcc cagcgccgtg 1200 taccgcaaga gcggcaccgt ggacagcctg gacgagatcc cccctcagaa caacaacgtg 1260 ccacctcgac agggcttcag ccaccgtctg agccacgtga gcatgttccg cagtggcttc 1320 agcaacagca gcgtgagcat catccgtgca cccatgttca gctggattca ccgcagcgcc 1380 accctgacca acaccatcga ccccgagcgc atcaaccaga tccccctggt gaagggcttc 1440 cgggtgtggg gcggcaccag cgtgatcacc ggccccggct tcaccggagg cgacatcctg 1500 cgcagaaaca ccttcggcga cttcgtgagc ctgcaggtga acatcaacag ccccatcacc 1560 cagcgttacc gcctgcgctt ccgctacgcc agcagccgcg acgcccgtgt gatcgtgctg 1620 actggcgccg ctagcaccgg tgtgggcggt caggtgagcg tgaacatgcc cctgcagaag 1680 actatggaga tcggcgagaa cctgactagt cgcaccttcc gctacaccga cttcagcaac 1740 cccttcagct tccgcgccaa ccccgacatc atcggcatca gcgagcagcc cctgttcggt 1800 gccggcagca tcagcagcgg cgagctgtac atcgacaaga tcgagatcat cctggccgac 1860 gccaccttcg aggccgagag cgacctggag cgcgcccaga aggccgtgaa cgccctgttc 1920 accagcagca accagatcgg cctgaagacc gacgtgaccg actaccacat cgaccaggtg 1980 agcaacctgg tggactgctt aagcgacgag ttctgcctgg acgagaagaa ggagctgagc 2040 gagaaggtga agcacgccaa gcgcctgagc gacgagcgca acctgctgca ggaccccaac 2100 ttccgcggca tcaaccgcca gctggaccgc ggctggcgag gcagcaccga tatcaccatc 2160 cagggcggcg acgacgtgtt caaggagaac tacgtgaccc tgcagggcac cttcgacgag 2220 tgctacccca cctacctgta ccagccgatc gacgagagca agctgaaggc ctacacccgc 2280 taccagctgc gcggctacat cgaggacagc caggacctgg aaatctacct gatccgctac 2340 aacgcgaagc acgagaccgt gaacgtgccc ggcaccggca gcctgtggcc cctgagcgcc 2400 cccagcccca tcggcaagtg cggggagccg aatcgatgcg ctccgcacct ggagtggaac 2460 ccggacctag actgcagctg cagggacggg gagaagtgcg cccaccacag ccaccacttc 2520 agcctggaca tcgacgtggg ctgcaccgac ctgaacgagg acctgggcgt gtgggtgatc 2580 ttcaagatca agacccagga cggccacgcc cgcctgggca atctagagtt cctggaggag 2640 aagcccctgg tgggcgaggc cctggcccgc gtgaagcgtg ctgagaagaa gtggcgcgac 2700 aagcgcgaga agctggagtg ggagaccaac atcgtgtaca aggaggccaa ggagagcgtg 2760 gacgccctgt tcgtgaacag ccagtacgac cgcctgcagg ccgacaccaa catcgccatg 2820 atccacgccg ccgacaagcg cgtgcacagc attcgcgagg cctacctgcc cgagctgagc 2880 gtgatccccg gtgtgaacgc cgccatcttc gaggaactcg agggccgcat cttcaccgcc 2940 ttcagcctgt acgacgcccg caacgtgatc aagaacggcg acttcaacaa cggcctgagc 3000 tgctggaacg tgaagggcca cgtggacgtg gaggagcaga acaaccaccg cagcgtgctg 3060 gtggtgcccg agtgggaggc cgaggtgagc caggaggtgc gcgtgtgccc cggccgcggc 3120 tacatcctgc gcgtgaccgc ctacaaggag ggctacggcg agggctgcgt gaccatccac 3180 gagatcgaga acaacaccga cgagctcaag ttcagcaact gcgtggagga ggaggtttac 3240 cccaacaaca ccgtgacctg caacgactac accgcgaccc aggaggagta cgaaggcacc 3300 tacacctctc gcaacagggg ttacgacggc gcctacgagt ccaacagctc cgtgccagct 3360 gactacgcca gcgcctacga ggagaaagcc tacaccgacg gtagacgcga caacccatgt 3420 gagagcaaca gaggctacgg cgactacacc cccctgcccg ctggatacgt gaccaaggag 3480 ctggagtact tccccgagac cgacaaggtg tggatcgaga ttggcgagac cgagggcacc 3540 ttcatcgtgg acagcgtgga gctgctgctg atggaggagt agtagatctg ttctgcacaa 3600 agtggagtag tcagtcatcg atcaggaacc agacaccaga cttttattca tacagtgaag 3660 tgaagtgaag tgcagtgcag tgagttgctg gtttttgtac aacttagtat gtatttgtat 3720 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc aggggtacca 3780 gcttgcatgc ctgcagtgca gcgtgacccg gtcgtgcccc tctctagaga taatgagcat 3840 tgcatgtcta agttataaaa aattaccaca tatttttttt gtcacacttg tttgaagtgc 3900 agtttatcta tctttataca tatatttaaa ctttactcta cgaataatat aatctatagt 3960 actacaataa tatcagtgtt ttagagaatc atataaatga acagttagac atggtctaaa 4020 ggacaattga gtattttgac aacaggactc tacagtttta tctttttagt gtgcatgtgt 4080 tctccttttt ttttgcaaat agcttcacct atataatact tcatccattt tattagtaca 4140 tccatttagg gtttagggtt aatggttttt atagactaat ttttttagta catctatttt 4200 attctatttt agcctctaaa ttaagaaaac taaaactcta ttttagtttt tttatttaat 4260 aatttagata taaaatagaa taaaataaag tgactaaaaa ttaaacaaat accctttaag 4320 aaattaaaaa aactaaggaa acatttttct tgtttcgagt agataatgcc agcctgttaa 4380 acgccgtcga cgagtctaac ggacaccaac cagcgaacca gcagcgtcgc gtcgggccaa 4440 gcgaagcaga cggcacggca tctctgtcgc tgcctctgga cccctctcga gagttccgct 4500 ccaccgttgg acttgctccg ctgtcggcat ccagaaattg cgtggcggag cggcagacgt 4560 gagccggcac ggcaggcggc ctcctcctcc tctcacggca ccggcagcta cgggggattc 4620 ctttcccacc gctccttcgc tttcccttcc tcgcccgccg taataaatag acaccccctc 4680 cacaccctct ttccccaacc tcgtgttgtt cggagcgcac acacacacaa ccagatctcc 4740 cccaaatcca cccgtcggca cctccgcttc aaggtacgcc gctcgtcctc cccccccccc 4800 cctctctacc ttctctagat cggcgttccg gtccatggtt agggcccggt agttctactt 4860 ctgttcatgt ttgtgttaga tccgtgtttg tgttagatcc gtgctgctag cgttcgtaca 4920 cggatgcgac ctgtacgtca gacacgttct gattgctaac ttgccagtgt ttctctttgg 4980 ggaatcctgg gatggctcta gccgttccgc agacgggatc gatttcatga ttttttttgt 5040 ttcgttgcat agggtttggt ttgccctttt cctttatttc aatatatgcc gtgcacttgt 5100 ttgtcgggtc atcttttcat gctttttttt gtcttggttg tgatgatgtg gtctggttgg 5160 gcggtcgttc tagatcggag tagaattctg tttcaaacta cctggtggat ttattaattt 5220 tggatctgta tgtgtgtgcc atacatattc atagttacga attgaagatg atggatggaa 5280 atatcgatct aggataggta tacatgttga tgcgggtttt actgatgcat atacagagat 5340 gctttttgtt cgcttggttg tgatgatgtg gtgtggttgg gcggtcgttc attcgttcta 5400 gatcggagta gaatactgtt tcaaactacc tggtgtattt attaattttg gaactgtatg 5460 tgtgtgtcat acatcttcat agttacgagt ttaagatgga tggaaatatc gatctaggat 5520 aggtatacat gttgatgtgg gttttactga tgcatataca tgatggcata tgcagcatct 5580 attcatatgc tctaaccttg agtacctatc tattataata aacaagtatg ttttataatt 5640 attttgatct tgatatactt ggatgatggc atatgcagca gctatatgtg gattttttta 5700 gccctgcctt catacgctat ttatttgctt ggtactgttt cttttgtcga tgctcaccct 5760 gttgtttggt gttacttctg cagggatccc cgatcatgca aaaactcatt aactcagtgc 5820 aaaactatgc ctggggcagc aaaacggcgt tgactgaact ttatggtatg gaaaatccgt 5880 ccagccagcc gatggccgag ctgtggatgg gcgcacatcc gaaaagcagt tcacgagtgc 5940 agaatgccgc cggagatatc gtttcactgc gtgatgtgat tgagagtgat aaatcgactc 6000 tgctcggaga ggccgttgcc aaacgctttg gcgaactgcc tttcctgttc aaagtattat 6060 gcgcagcaca gccactctcc attcaggttc atccaaacaa acacaattct gaaatcggtt 6120 ttgccaaaga aaatgccgca ggtatcccga tggatgccgc cgagcgtaac tataaagatc 6180 ctaaccacaa gccggagctg gtttttgcgc tgacgccttt ccttgcgatg aacgcgtttc 6240 gtgaattttc cgagattgtc tccctactcc agccggtcgc aggtgcacat ccggcgattg 6300 ctcacttttt acaacagcct gatgccgaac gtttaagcga actgttcgcc agcctgttga 6360 atatgcaggg tgaagaaaaa tcccgcgcgc tggcgatttt aaaatcggcc ctcgatagcc 6420 agcagggtga accgtggcaa acgattcgtt taatttctga attttacccg gaagacagcg 6480 gtctgttctc cccgctattg ctgaatgtgg tgaaattgaa ccctggcgaa gcgatgttcc 6540 tgttcgctga aacaccgcac gcttacctgc aaggcgtggc gctggaagtg atggcaaact 6600 ccgataacgt gctgcgtgcg ggtctgacgc ctaaatacat tgatattccg gaactggttg 6660 ccaatgtgaa attcgaagcc aaaccggcta accagttgtt gacccagccg gtgaaacaag 6720 gtgcagaact ggacttcccg attccagtgg atgattttgc cttctcgctg catgacctta 6780 gtgataaaga aaccaccatt agccagcaga gtgccgccat tttgttctgc gtcgaaggcg 6840 atgcaacgtt gtggaaaggt tctcagcagt tacagcttaa accgggtgaa tcagcgttta 6900 ttgccgccaa cgaatcaccg gtgactgtca aaggccacgg ccgtttagcg cgtgtttaca 6960 acaagctgta agagcttact gaaaaaatta acatctcttg ctaagctggg agctcgatcc 7020 gtcgacctgc agatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg 7080 ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta 7140 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat 7200 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg 7260 cggtgtcatc tatgttacta gatctgctag ccctgcagga aatttaccgg tgcccgggcg 7320 gccagcatgg ccgtatccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac 7380 cacaatatat cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat 7440 caccactcga tacaggcagc ccatcagaat taattctcat gtttgacagc ttatcatcga 7500 ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt 7560 gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat 7620 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat 7680 taatcatccg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa 7740 cagaccatga gggaagcgtt gatcgccgaa gtatcgactc aactatcaga ggtagttggc 7800 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg 7860 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt 7920 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga 7980 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg 8040 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt 8100 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca 8160 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct 8220 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac 8280 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta 8340 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc 8400 cagtatcagc ccgtcatact tgaagctagg caggcttatc ttggacaaga agatcgcttg 8460 gcctcgcgcg cagatcagtt ggaagaattt gttcactacg tgaaaggcga gatcaccaaa 8520 gtagtcggca aataaagctc tagtggatct ccgtaccccc gggggatctg gctcgcggcg 8580 gacgcacgac gccggggcga gaccataggc gatctcctaa atcaatagta gctgtaacct 8640 cgaagcgttt cacttgtaac aacgattgag aatttttgtc ataaaattga aatacttggt 8700 tcgcattttt gtcatccgcg gtcagccgca attctgacga actgcccatt tagctggaga 8760 tgattgtaca tccttcacgt gaaaatttct caagcgctgt gaacaagggt tcagatttta 8820 gattgaaagg tgagccgttg aaacacgttc ttcttgtcga tgacgacgtc gctatgcggc 8880 atcttattat tgaatacctt acgatccacg ccttcaaagt gaccgcggta gccgacagca 8940 cccagttcac aagagtactc tcttccgcga cggtcgatgt cgtggttgtt gatctaaatt 9000 taggtcgtga agatgggctc gagatcgttc gtaatctggc ggcaaagtct gatattccaa 9060 tcataattat cagtggcgac cgccttgagg agacggataa agttgttgca ctcgagctag 9120 gagcaagtga ttttatcgct aagccgttca gtatcagaga gtttctagca cgcattcggg 9180 ttgccttgcg cgtgcgcccc aacgttgtcc gctccaaaga ccgacggtct ttttgtttta 9240 ctgactggac acttaatctc aggcaacgtc gcttgatgtc cgaagctggc ggtgaggtga 9300 aacttacggc aggtgagttc aatcttctcc tcgcgttttt agagaaaccc cgcgacgttc 9360 tatcgcgcga gcaacttctc attgccagtc gagtacgcga cgaggaggtt tatgacagga 9420 gtatagatgt tctcattttg aggctgcgcc gcaaacttga ggcagatccg tcaagccctc 9480 aactgataaa aacagcaaga ggtgccggtt atttctttga cgcggacgtg caggtttcgc 9540 acggggggac gatggcagcc tgagccaatt cccagatccc cgaggaatcg gcgtgagcgg 9600 tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa 9660 gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga 9720 atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg 9780 tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat 9840 gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct 9900 gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt 9960 agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat 10020 ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc 10080 cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg 10140 cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc 10200 catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc 10260 cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat 10320 cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg acgtgctgac 10380 ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc 10440 acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag 10500 tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa 10560 tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat 10620 gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat 10680 gctagggcaa attgccctag caggggaaaa aggtcgaaaa ggtctctttc ctgtggatag 10740 cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc 10800 aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg 10860 cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg 10920 tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta cccttcggtc 10980 gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa 11040 aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc cgtcgccact 11100 cgaccgccgg cgctgaggtc tgcctcgtga agaaggtgtt gctgactcat accaggcctg 11160 aatcgcccca tcatccagcc agaaagtgag ggagccacgg ttgatgagag ctttgttgta 11220 ggtggaccag ttggtgattt tgaacttttg ctttgccacg gaacggtctg cgttgtcggg 11280 aagatgcgtg atctgatcct tcaactcagc aaaagttcga tttattcaac aaagccgccg 11340 tcccgtcaag tcagcgtaat gctctgccag tgttacaacc aattaaccaa ttctgattag 11400 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca 11460 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg 11520 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt 11580 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa 11640 tccggtgaga atggcaaaag ctctgcatta atgaatcggc caacgcgcgg ggagaggcgg 11700 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 11760 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 11820 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 11880 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 11940 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 12000 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 12060 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 12120 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 12180 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 12240 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 12300 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 12360 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 12420 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 12480 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 12540 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttgat 12600 ccggaattaa ttcctgtggt tggcatgcac atacaaatgg acgaacggat aaaccttttc 12660 acgccctttt aaatatccga ttattctaat aaacgctctt ttctcttagg tttacccgcc 12720 aatatatcct gtcaaacact gatagtttaa actgaaggcg ggaaacgaca atctgatcat 12780 gagcggagaa ttaagggagt cacgttatga cccccgccga tgacgcggga caagccgttt 12840 tacgtttgga actgacagaa ccgcaacgct gcaggaattg gccgcagcgg ccatttaaat 12900 caattgggcg cgccgaattc gagctcggta caagcttgca catgacaaca attgtaagag 12960 gatggagacc acaacgatcc aacaatactt ctgcgacggg ctgtgaagta tagagaagtt 13020 aaacgcccaa aagccattgt gtttggaatt tttagttatt ctatttttca tgatgtatct 13080 tcctctaaca tgccttaatt tgcaaatttg gtataactac tgattgaaaa tatatgtatg 13140 taaaaaaata ctaagcatat ttgtgaagct aaacatgatg ttatttaaga aaatatgttg 13200 ttaacagaat aagattaata tcgaaatgga aacatctgta aattagaatc atcttacaag 13260 ctaagagatg ttcacgcttt gagaaacttc ttcagatcat gaccgtagaa gtagctctcc 13320 aagactcaac gaaggctgct gcaattccac aaatgcatga catgcatcct tgtaaccgtc 13380 gtcgccgcta taaacacgga taactcaatt ccctgctcca tcaatttaga aatgagcaag 13440 caagcacccg atcgctcacc ccatatgcac caatctgact cccaagtctc tgtttcgcat 13500 tagtaccgcc agcactccac ctatagctac caattgagac ctttccagcc taagcagatc 13560 gattgatcgt tagagtcaaa gagttggtgg tacgggtact ttaactacca tggaatgatg 13620 gggcgtgatg tagagcggaa agcgcctccc tacgcggaac aacaccctcg ccatgccgct 13680 cgactacagc ctcctcctcg tcggccgccc acaacgaggg agcccgtggt cgcagccacc 13740 gaccagcatg tctctgtgtc ctcgtccgac ctcgacatgt catggcaaac agtcggacgc 13800 cagcaccaga ctgacgacat gagtctctga agagcccgcc acctagaaag atccgagccc 13860 tgctgctggt agtggtaacc attttcgtcg cgctgacgcg gagagcgaga ggccagaaat 13920 ttatagcgac tgacgctgtg gcaggcacgc tatcggaggt tacgacgtgg cgggtcactc 13980 gacgcggagt tcacaggtcc tatccttgca tcgctcgggc cggagtttac gggacttatc 14040 cttacgacgt gctctaaggt tgcgataacg ggcggaggaa ggcgtgtggc gtgcggagac 14100 ggtttataca cgtagtgtgc gggagtgtgt ttcgtagacg cgggaaagca cgacgactta 14160 cgaaggttag tggaggagga ggacacacta aaatcaggac gcaagaaact cttctattat 14220 agtagtagag aagagattat aggagtgtgg gttgattcta aagaaaatcg acgcaggaca 14280 accgtcaaaa cgggtgcttt aatatagtag atatatatat atagagagag agagaaagta 14340 caaaggatgc atttgtgtct gcatatgatc ggagtattac taacggccgt cgtaagaagg 14400 tccatcatgc gtggagcgag cccatttggt tggttgtcag gccgcagtta aggcctccat 14460 atatgattgt cgtcgggccc ataacagcat ctcctccacc agtttattgt aagaataaat 14520 taagtagaga tatttgtcgt cgggcagaag aaacttggac aagaagaaga agcaagctag 14580 gccaatttct tgccggcaag aggaagatag tggcctctag tttatatatc ggcgtgatga 14640 tgatgctcct agctagaaat gagagaagaa aaacggacgc gtgtttggtg tgtgtcaatg 14700 gcgtccatcc ttccatcaga tcagaacgat gaaaaagtca agcacggcat gcatagtata 14760 tgtatagctt gttttagtgt ggctttgctg agacgaatga aagcaacggc gggcatattt 14820 ttcagtggct gtagctttca ggctgaaaga gacgtggcat gcaataattc agggaattcg 14880 tcagccaatt gaggtagcta gtcaacttgt acattggtgc gagcaatttt ccgcactcag 14940 gagggctagt ttgagagtcc aaaaactata ggagattaaa gaggctaaaa tcctctcctt 15000 atttaatttt aaataagtag tgtatttgta ttttaactcc tccaaccctt ccgattttat 15060 ggctctcaaa ctagcattca gtctaatgca tgcatgcttg gctagaggtc gtatggggtt 15120 gttaatagca tagctagcta caagttaacc gggtctttta tatttaataa ggacaggcaa 15180 agtattactt acaaataaag aataaagcta ggacgaactc gtggattatt actaaatcga 15240 aatggacgta atattccagg caagaataat tgttcgatca ggagacaagt ggggcattgg 15300 accggttctt gcaagcaaga gcctatggcg tggtgacacg gcgcgttgcc catacatcat 15360 gcctccatcg atgatccatc ctcacttgct ataaaaagag gtgtccatgg tgctcaagct 15420 cagccaagca aataagacga cttgtttcat tgattcttca agagatcgag cttcttttgc 15480 accacaaggt cgaggatcca aca 15503 <210> 15 <211> 14946 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: pNOV1313 <220> <221> misc_feature (222) (12). (1993) <223> Zm Ubi promoter <220> <221> misc_feature <222> (2016) .. (5597) <223> synthetic nucleotide sequence encoding the toxin       portion of H04 plus a full-length Cry1Ab tail       portion <220> <221> misc_feature (222) (5805) .. (7786) <223> Zm Ubi promoter <220> <221> misc_feature (222) (7883) .. (8986) <223> PMI <400> 15 aagcttgcat gcctgcagtg cagcgtgacc cggtcgtgcc cctctctaga gataatgagc 60 attgcatgtc taagttataa aaaattacca catatttttt ttgtcacact tgtttgaagt 120 gcagtttatc tatctttata catatattta aactttactc tacgaataat ataatctata 180 gtactacaat aatatcagtg ttttagagaa tcatataaat gaacagttag acatggtcta 240 aaggacaatt gagtattttg acaacaggac tctacagttt tatcttttta gtgtgcatgt 300 gttctccttt ttttttgcaa atagcttcac ctatataata cttcatccat tttattagta 360 catccattta gggtttaggg ttaatggttt ttatagacta atttttttag tacatctatt 420 ttattctatt ttagcctcta aattaagaaa actaaaactc tattttagtt tttttattta 480 ataatttaga tataaaatag aataaaataa agtgactaaa aattaaacaa atacccttta 540 agaaattaaa aaaactaagg aaacattttt cttgtttcga gtagataatg ccagcctgtt 600 aaacgccgtc gacgagtcta acggacacca accagcgaac cagcagcgtc gcgtcgggcc 660 aagcgaagca gacggcacgg catctctgtc gctgcctctg gacccctctc gagagttccg 720 ctccaccgtt ggacttgctc cgctgtcggc atccagaaat tgcgtggcgg agcggcagac 780 gtgagccggc acggcaggcg gcctcctcct cctctcacgg caccggcagc tacgggggat 840 tcctttccca ccgctccttc gctttccctt cctcgcccgc cgtaataaat agacaccccc 900 tccacaccct ctttccccaa cctcgtgttg ttcggagcgc acacacacac aaccagatct 960 cccccaaatc cacccgtcgg cacctccgct tcaaggtacg ccgctcgtcc tccccccccc 1020 cccctctcta ccttctctag atcggcgttc cggtccatgg ttagggcccg gtagttctac 1080 ttctgttcat gtttgtgtta gatccgtgtt tgtgttagat ccgtgctgct agcgttcgta 1140 cacggatgcg acctgtacgt cagacacgtt ctgattgcta acttgccagt gtttctcttt 1200 ggggaatcct gggatggctc tagccgttcc gcagacggga tcgatttcat gatttttttt 1260 gtttcgttgc atagggtttg gtttgccctt ttcctttatt tcaatatatg ccgtgcactt 1320 gtttgtcggg tcatcttttc atgctttttt ttgtcttggt tgtgatgatg tggtctggtt 1380 gggcggtcgt tctagatcgg agtagaattc tgtttcaaac tacctggtgg atttattaat 1440 tttggatctg tatgtgtgtg ccatacatat tcatagttac gaattgaaga tgatggatgg 1500 aaatatcgat ctaggatagg tatacatgtt gatgcgggtt ttactgatgc atatacagag 1560 atgctttttg ttcgcttggt tgtgatgatg tggtgtggtt gggcggtcgt tcattcgttc 1620 tagatcggag tagaatactg tttcaaacta cctggtgtat ttattaattt tggaactgta 1680 tgtgtgtgtc atacatcttc atagttacga gtttaagatg gatggaaata tcgatctagg 1740 ataggtatac atgttgatgt gggttttact gatgcatata catgatggca tatgcagcat 1800 ctattcatat gctctaacct tgagtaccta tctattataa taaacaagta tgttttataa 1860 ttattttgat cttgatatac ttggatgatg gcatatgcag cagctatatg tggatttttt 1920 tagccctgcc ttcatacgct atttatttgc ttggtactgt ttcttttgtc gatgctcacc 1980 ctgttgtttg gtgttacttc tgcagggatc caacaatgga caacaacccc aacatcaacg 2040 agtgcatccc ctacaactgc ctgagcaacc ccgaggtgga ggtgctgggc ggcgagcgca 2100 tcgagaccgg ctacaccccc atcgacatca gcctgagcct gacccagttc ctgctgagcg 2160 agttcgtgcc cggcgccggc ttcgtgctgg gcctggtgga catcatctgg ggcatcttcg 2220 gccccagcca gtgggacgcc ttcctggtgc agatcgagca gttgataaac caacgcatag 2280 aggaattcgc ccgcaaccag gccatcagcc gcctggaggg cctgagcaac ctgtaccaaa 2340 tctacgccga gagcttccgc gagtgggagg ccgaccccac caaccccgcc ctgcgcgagg 2400 agatgcgcat ccagttcaac gacatgaaca gcgccctgac caccgccatc cccctgttcg 2460 ccgtgcagaa ctaccaggtg cccctgctga gcgtgtacgt gcaggccgcc aacctgcacc 2520 tgagcgtgct gcgcgacgtc agcgtgttcg gccagcgctg gggcttcgac gccgccacca 2580 tcaacagccg ctacaacgac ctgacccgcc tgatcggcaa ctacaccgac cacgccgtgc 2640 gctggtacaa caccggcctg gagcgcgtgt ggggtcccga cagccgcgac tggatcaggt 2700 acaaccagtt ccgccgcgag ctgaccctga ccgtgctgga catcgtgagc ctgttcccca 2760 actacgacag ccgcacctac cccatccgca ccgtgagcca gctgacccgc gagatttaca 2820 ccaaccccgt gctggagaac ttcgacggca gcttccgcgg cagcgcccag ggcatcgagg 2880 gcagcatccg cagcccccac ctgatggaca tcctgaacag catcaccatc tacaccgacg 2940 cccaccgcgg cgagtactac tggagcggcc accagatcat ggccagcccc gtcggcttca 3000 gcggccccga gttcaccttc cccctgtacg gcaccatggg caacgctgca cctcagcagc 3060 gcatcgtggc acagctgggc cagggagtgt accgcaccct gagcagcacc ctgtaccgtc 3120 gacctttcaa catcggcatc aacaaccagc agctgagcgt gctggacggc accgagttcg 3180 cctacggcac cagcagcaac ctgcccagcg ccgtgtaccg caagagcggc accgtggaca 3240 gcctggacga gatcccccct cagaacaaca acgtgccacc tcgacagggc ttcagccacc 3300 gtctgagcca cgtgagcatg ttccgcagtg gcttcagcaa cagcagcgtg agcatcatcc 3360 gtgcacccat gttcagctgg attcaccgca gcgccaccct gaccaacacc atcgaccccg 3420 agcgcatcaa ccagatcccc ctggtgaagg gcttccgggt gtggggcggc accagcgtga 3480 tcaccggccc cggcttcacc ggaggcgaca tcctgcgcag aaacaccttc ggcgacttcg 3540 tgagcctgca ggtgaacatc aacagcccca tcacccagcg ttaccgcctg cgcttccgct 3600 acgccagcag ccgcgacgcc cgtgtgatcg tgctgactgg cgccgctagc accggtgtgg 3660 gcggtcaggt gagcgtgaac atgcccctgc agaagactat ggagatcggc gagaacctga 3720 ctagtcgcac cttccgctac accgacttca gcaacccctt cagcttccgc gccaaccccg 3780 acatcatcgg catcagcgag cagcccctgt tcggtgccgg cagcatcagc agcggcgagc 3840 tgtacatcga caagatcgag atcatcctgg ccgacgccac cttcgaggcc gagagcgacc 3900 tggagcgcgc ccagaaggcc gtgaacgccc tgttcaccag cagcaaccag atcggcctga 3960 agaccgacgt gaccgactac cacatcgacc aggtgagcaa cctggtggac tgcttaagcg 4020 acgagttctg cctggacgag aagaaggagc tgagcgagaa ggtgaagcac gccaagcgcc 4080 tgagcgacga gcgcaacctg ctgcaggacc ccaacttccg cggcatcaac cgccagctgg 4140 accgcggctg gcgaggcagc accgatatca ccatccaggg cggcgacgac gtgttcaagg 4200 agaactacgt gaccctgcag ggcaccttcg acgagtgcta ccccacctac ctgtaccagc 4260 cgatcgacga gagcaagctg aaggcctaca cccgctacca gctgcgcggc tacatcgagg 4320 acagccagga cctggaaatc tacctgatcc gctacaacgc gaagcacgag accgtgaacg 4380 tgcccggcac cggcagcctg tggcccctga gcgcccccag ccccatcggc aagtgcgggg 4440 agccgaatcg atgcgctccg cacctggagt ggaacccgga cctagactgc agctgcaggg 4500 acggggagaa gtgcgcccac cacagccacc acttcagcct ggacatcgac gtgggctgca 4560 ccgacctgaa cgaggacctg ggcgtgtggg tgatcttcaa gatcaagacc caggacggcc 4620 acgcccgcct gggcaatcta gagttcctgg aggagaagcc cctggtgggc gaggccctgg 4680 cccgcgtgaa gcgtgctgag aagaagtggc gcgacaagcg cgagaagctg gagtgggaga 4740 ccaacatcgt gtacaaggag gccaaggaga gcgtggacgc cctgttcgtg aacagccagt 4800 acgaccgcct gcaggccgac accaacatcg ccatgatcca cgccgccgac aagcgcgtgc 4860 acagcattcg cgaggcctac ctgcccgagc tgagcgtgat ccccggtgtg aacgccgcca 4920 tcttcgagga actcgagggc cgcatcttca ccgccttcag cctgtacgac gcccgcaacg 4980 tgatcaagaa cggcgacttc aacaacggcc tgagctgctg gaacgtgaag ggccacgtgg 5040 acgtggagga gcagaacaac caccgcagcg tgctggtggt gcccgagtgg gaggccgagg 5100 tgagccagga ggtgcgcgtg tgccccggcc gcggctacat cctgcgcgtg accgcctaca 5160 aggagggcta cggcgagggc tgcgtgacca tccacgagat cgagaacaac accgacgagc 5220 tcaagttcag caactgcgtg gaggaggagg tttaccccaa caacaccgtg acctgcaacg 5280 actacaccgc gacccaggag gagtacgaag gcacctacac ctctcgcaac aggggttacg 5340 acggcgccta cgagtccaac agctccgtgc cagctgacta cgccagcgcc tacgaggaga 5400 aagcctacac cgacggtaga cgcgacaacc catgtgagag caacagaggc tacggcgact 5460 acacccccct gcccgctgga tacgtgacca aggagctgga gtacttcccc gagaccgaca 5520 aggtgtggat cgagattggc gagaccgagg gcaccttcat cgtggacagc gtggagctgc 5580 tgctgatgga ggagtagtag atctgttctg cacaaagtgg agtagtcagt catcgatcag 5640 gaaccagaca ccagactttt attcatacag tgaagtgaag tgaagtgcag tgcagtgagt 5700 tgctggtttt tgtacaactt agtatgtatt tgtatttgta aaatacttct atcaataaaa 5760 tttctaattc ctaaaaccaa aatccagggg taccagcttg catgcctgca gtgcagcgtg 5820 acccggtcgt gcccctctct agagataatg agcattgcat gtctaagtta taaaaaatta 5880 ccacatattt tttttgtcac acttgtttga agtgcagttt atctatcttt atacatatat 5940 ttaaacttta ctctacgaat aatataatct atagtactac aataatatca gtgttttaga 6000 gaatcatata aatgaacagt tagacatggt ctaaaggaca attgagtatt ttgacaacag 6060 gactctacag ttttatcttt ttagtgtgca tgtgttctcc tttttttttg caaatagctt 6120 cacctatata atacttcatc cattttatta gtacatccat ttagggttta gggttaatgg 6180 tttttataga ctaatttttt tagtacatct attttattct attttagcct ctaaattaag 6240 aaaactaaaa ctctatttta gtttttttat ttaataattt agatataaaa tagaataaaa 6300 taaagtgact aaaaattaaa caaataccct ttaagaaatt aaaaaaacta aggaaacatt 6360 tttcttgttt cgagtagata atgccagcct gttaaacgcc gtcgacgagt ctaacggaca 6420 ccaaccagcg aaccagcagc gtcgcgtcgg gccaagcgaa gcagacggca cggcatctct 6480 gtcgctgcct ctggacccct ctcgagagtt ccgctccacc gttggacttg ctccgctgtc 6540 ggcatccaga aattgcgtgg cggagcggca gacgtgagcc ggcacggcag gcggcctcct 6600 cctcctctca cggcaccggc agctacgggg gattcctttc ccaccgctcc ttcgctttcc 6660 cttcctcgcc cgccgtaata aatagacacc ccctccacac cctctttccc caacctcgtg 6720 ttgttcggag cgcacacaca cacaaccaga tctcccccaa atccacccgt cggcacctcc 6780 gcttcaaggt acgccgctcg tcctcccccc ccccccctct ctaccttctc tagatcggcg 6840 ttccggtcca tggttagggc ccggtagttc tacttctgtt catgtttgtg ttagatccgt 6900 gtttgtgtta gatccgtgct gctagcgttc gtacacggat gcgacctgta cgtcagacac 6960 gttctgattg ctaacttgcc agtgtttctc tttggggaat cctgggatgg ctctagccgt 7020 tccgcagacg ggatcgattt catgattttt tttgtttcgt tgcatagggt ttggtttgcc 7080 cttttccttt atttcaatat atgccgtgca cttgtttgtc gggtcatctt ttcatgcttt 7140 tttttgtctt ggttgtgatg atgtggtctg gttgggcggt cgttctagat cggagtagaa 7200 ttctgtttca aactacctgg tggatttatt aattttggat ctgtatgtgt gtgccataca 7260 tattcatagt tacgaattga agatgatgga tggaaatatc gatctaggat aggtatacat 7320 gttgatgcgg gttttactga tgcatataca gagatgcttt ttgttcgctt ggttgtgatg 7380 atgtggtgtg gttgggcggt cgttcattcg ttctagatcg gagtagaata ctgtttcaaa 7440 ctacctggtg tatttattaa ttttggaact gtatgtgtgt gtcatacatc ttcatagtta 7500 cgagtttaag atggatggaa atatcgatct aggataggta tacatgttga tgtgggtttt 7560 actgatgcat atacatgatg gcatatgcag catctattca tatgctctaa ccttgagtac 7620 ctatctatta taataaacaa gtatgtttta taattatttt gatcttgata tacttggatg 7680 atggcatatg cagcagctat atgtggattt ttttagccct gccttcatac gctatttatt 7740 tgcttggtac tgtttctttt gtcgatgctc accctgttgt ttggtgttac ttctgcaggg 7800 atccccgatc atgcaaaaac tcattaactc agtgcaaaac tatgcctggg gcagcaaaac 7860 ggcgttgact gaactttatg gtatggaaaa tccgtccagc cagccgatgg ccgagctgtg 7920 gatgggcgca catccgaaaa gcagttcacg agtgcagaat gccgccggag atatcgtttc 7980 actgcgtgat gtgattgaga gtgataaatc gactctgctc ggagaggccg ttgccaaacg 8040 ctttggcgaa ctgcctttcc tgttcaaagt attatgcgca gcacagccac tctccattca 8100 ggttcatcca aacaaacaca attctgaaat cggttttgcc aaagaaaatg ccgcaggtat 8160 cccgatggat gccgccgagc gtaactataa agatcctaac cacaagccgg agctggtttt 8220 tgcgctgacg cctttccttg cgatgaacgc gtttcgtgaa ttttccgaga ttgtctccct 8280 actccagccg gtcgcaggtg cacatccggc gattgctcac tttttacaac agcctgatgc 8340 cgaacgttta agcgaactgt tcgccagcct gttgaatatg cagggtgaag aaaaatcccg 8400 cgcgctggcg attttaaaat cggccctcga tagccagcag ggtgaaccgt ggcaaacgat 8460 tcgtttaatt tctgaatttt acccggaaga cagcggtctg ttctccccgc tattgctgaa 8520 tgtggtgaaa ttgaaccctg gcgaagcgat gttcctgttc gctgaaacac cgcacgctta 8580 cctgcaaggc gtggcgctgg aagtgatggc aaactccgat aacgtgctgc gtgcgggtct 8640 gacgcctaaa tacattgata ttccggaact ggttgccaat gtgaaattcg aagccaaacc 8700 ggctaaccag ttgttgaccc agccggtgaa acaaggtgca gaactggact tcccgattcc 8760 agtggatgat tttgccttct cgctgcatga ccttagtgat aaagaaacca ccattagcca 8820 gcagagtgcc gccattttgt tctgcgtcga aggcgatgca acgttgtgga aaggttctca 8880 gcagttacag cttaaaccgg gtgaatcagc gtttattgcc gccaacgaat caccggtgac 8940 tgtcaaaggc cacggccgtt tagcgcgtgt ttacaacaag ctgtaagagc ttactgaaaa 9000 aattaacatc tcttgctaag ctgggagctc gatccgtcga cctgcagatc gttcaaacat 9060 ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata 9120 atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat 9180 gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa 9240 aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatct 9300 gctagccctg caggaaattt accggtgccc gggcggccag catggccgta tccgcaatgt 9360 gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag 9420 ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc 9480 agaattaatt ctcatgtttg acagcttatc atcgactgca cggtgcacca atgcttctgg 9540 cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt 9600 cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg 9660 ttctggcaaa tattctgaaa tgagctgttg acaattaatc atccggctcg tataatgtgt 9720 ggaattgtga gcggataaca atttcacaca ggaaacagac catgagggaa gcgttgatcg 9780 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga 9840 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg 9900 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga 9960 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag 10020 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac 10080 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga 10140 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag 10200 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa 10260 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag 10320 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg 10380 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag 10440 ctaggcaggc ttatcttgga caagaagatc gcttggcctc gcgcgcagat cagttggaag 10500 aatttgttca ctacgtgaaa ggcgagatca ccaaagtagt cggcaaataa agctctagtg 10560 gatctccgta cccccggggg atctggctcg cggcggacgc acgacgccgg ggcgagacca 10620 taggcgatct cctaaatcaa tagtagctgt aacctcgaag cgtttcactt gtaacaacga 10680 ttgagaattt ttgtcataaa attgaaatac ttggttcgca tttttgtcat ccgcggtcag 10740 ccgcaattct gacgaactgc ccatttagct ggagatgatt gtacatcctt cacgtgaaaa 10800 tttctcaagc gctgtgaaca agggttcaga ttttagattg aaaggtgagc cgttgaaaca 10860 cgttcttctt gtcgatgacg acgtcgctat gcggcatctt attattgaat accttacgat 10920 ccacgccttc aaagtgaccg cggtagccga cagcacccag ttcacaagag tactctcttc 10980 cgcgacggtc gatgtcgtgg ttgttgatct aaatttaggt cgtgaagatg ggctcgagat 11040 cgttcgtaat ctggcggcaa agtctgatat tccaatcata attatcagtg gcgaccgcct 11100 tgaggagacg gataaagttg ttgcactcga gctaggagca agtgatttta tcgctaagcc 11160 gttcagtatc agagagtttc tagcacgcat tcgggttgcc ttgcgcgtgc gccccaacgt 11220 tgtccgctcc aaagaccgac ggtctttttg ttttactgac tggacactta atctcaggca 11280 acgtcgcttg atgtccgaag ctggcggtga ggtgaaactt acggcaggtg agttcaatct 11340 tctcctcgcg tttttagaga aaccccgcga cgttctatcg cgcgagcaac ttctcattgc 11400 cagtcgagta cgcgacgagg aggtttatga caggagtata gatgttctca ttttgaggct 11460 gcgccgcaaa cttgaggcag atccgtcaag ccctcaactg ataaaaacag caagaggtgc 11520 cggttatttc tttgacgcgg acgtgcaggt ttcgcacggg gggacgatgg cagcctgagc 11580 caattcccag atccccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa 11640 atcggcgcgg cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag 11700 cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga 11760 atccgcaaag aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc 11820 aagggcgacg agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat 11880 agtcgcagca tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc 11940 gaggtgatcc gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc 12000 atggccagtg tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc 12060 atgaaccgat accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt 12120 gcggacgtac tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta 12180 gaaacctgca ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag 12240 aacggccgcc tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta 12300 aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc 12360 gagatcacag aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc 12420 gatcccggca tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa 12480 gccagatggt tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag 12540 ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag 12600 gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc 12660 gaagcatccg ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg 12720 gaaaaaggtc gaaaaggtct ctttcctgtg gatagcacgt acattgggaa cccaaagccg 12780 tacattggga accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca 12840 cacatgtaag tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta 12900 aaacttatta aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca 12960 gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct 13020 tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat 13080 ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc gccggcgctg aggtctgcct 13140 cgtgaagaag gtgttgctga ctcataccag gcctgaatcg ccccatcatc cagccagaaa 13200 gtgagggagc cacggttgat gagagctttg ttgtaggtgg accagttggt gattttgaac 13260 ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac 13320 tcagcaaaag ttcgatttat tcaacaaagc cgccgtcccg tcaagtcagc gtaatgctct 13380 gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa 13440 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta 13500 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg 13560 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt 13620 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagctctg 13680 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 13740 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 13800 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 13860 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 13920 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 13980 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 14040 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 14100 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 14160 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 14220 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 14280 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 14340 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 14400 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 14460 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 14520 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 14580 ttatcaaaaa ggatcttcac ctagatcctt ttgatccgga attaattcct gtggttggca 14640 tgcacataca aatggacgaa cggataaacc ttttcacgcc cttttaaata tccgattatt 14700 ctaataaacg ctcttttctc ttaggtttac ccgccaatat atcctgtcaa acactgatag 14760 tttaaactga aggcgggaaa cgacaatctg atcatgagcg gagaattaag ggagtcacgt 14820 tatgaccccc gccgatgacg cgggacaagc cgttttacgt ttggaactga cagaaccgca 14880 acgctgcagg aattggccgc agcggccatt taaatcaatt gggcgcgccg aattcgagct 14940 cggtac 14946 <210> 16 <211> 14603 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: pNOV1435 <220> <221> misc_feature (222) (1) .. (2007) <223> synthetic nucleotide sequence encoding the toxin       portion of H04 plus the first 40 amino acids of       the Cry1Ab tail <220> <221> misc_feature (222) Complement ((8814) .. (10022)) <223> PMI <220> <221> misc_feature <222> (11142) .. (12032) <223> Maize ubiquitin promoter <220> <221> misc_feature (222) (12037) .. (14594) <223> MTL promoter <400> 16 atggacaaca accccaacat caacgagtgc atcccctaca actgcctgag caaccccgag 60 gtggaggtgc tgggcggcga gcgcatcgag accggctaca cccccatcga catcagcctg 120 agcctgaccc agttcctgct gagcgagttc gtgcccggcg ccggcttcgt gctgggcctg 180 gtggacatca tctggggcat cttcggcccc agccagtggg acgccttcct ggtgcagatc 240 gagcagttga taaaccaacg catagaggaa ttcgcccgca accaggccat cagccgcctg 300 gagggcctga gcaacctgta ccaaatctac gccgagagct tccgcgagtg ggaggccgac 360 cccaccaacc ccgccctgcg cgaggagatg cgcatccagt tcaacgacat gaacagcgcc 420 ctgaccaccg ccatccccct gttcgccgtg cagaactacc aggtgcccct gctgagcgtg 480 tacgtgcagg ccgccaacct gcacctgagc gtgctgcgcg acgtcagcgt gttcggccag 540 cgctggggct tcgacgccgc caccatcaac agccgctaca acgacctgac ccgcctgatc 600 ggcaactaca ccgaccacgc cgtgcgctgg tacaacaccg gcctggagcg cgtgtggggt 660 cccgacagcc gcgactggat caggtacaac cagttccgcc gcgagctgac cctgaccgtg 720 ctggacatcg tgagcctgtt ccccaactac gacagccgca cctaccccat ccgcaccgtg 780 agccagctga cccgcgagat ttacaccaac cccgtgctgg agaacttcga cggcagcttc 840 cgcggcagcg cccagggcat cgagggcagc atccgcagcc cccacctgat ggacatcctg 900 aacagcatca ccatctacac cgacgcccac cgcggcgagt actactggag cggccaccag 960 atcatggcca gccccgtcgg cttcagcggc cccgagttca ccttccccct gtacggcacc 1020 atgggcaacg ctgcacctca gcagcgcatc gtggcacagc tgggccaggg agtgtaccgc 1080 accctgagca gcaccctgta ccgtcgacct ttcaacatcg gcatcaacaa ccagcagctg 1140 agcgtgctgg acggcaccga gttcgcctac ggcaccagca gcaacctgcc cagcgccgtg 1200 taccgcaaga gcggcaccgt ggacagcctg gacgagatcc cccctcagaa caacaacgtg 1260 ccacctcgac agggcttcag ccaccgtctg agccacgtga gcatgttccg cagtggcttc 1320 agcaacagca gcgtgagcat catccgtgca cccatgttca gctggattca ccgcagcgcc 1380 accctgacca acaccatcga ccccgagcgc atcaaccaga tccccctggt gaagggcttc 1440 cgggtgtggg gcggcaccag cgtgatcacc ggccccggct tcaccggagg cgacatcctg 1500 cgcagaaaca ccttcggcga cttcgtgagc ctgcaggtga acatcaacag ccccatcacc 1560 cagcgttacc gcctgcgctt ccgctacgcc agcagccgcg acgcccgtgt gatcgtgctg 1620 actggcgccg ctagcaccgg tgtgggcggt caggtgagcg tgaacatgcc cctgcagaag 1680 actatggaga tcggcgagaa cctgactagt cgcaccttcc gctacaccga cttcagcaac 1740 cccttcagct tccgcgccaa ccccgacatc atcggcatca gcgagcagcc cctgttcggt 1800 gccggcagca tcagcagcgg cgagctgtac atcgacaaga tcgagatcat cctggccgac 1860 gccaccttcg aggccgagag cgacctggag cgcgcccaga aggccgtgaa cgccctgttc 1920 accagcagca accagatcgg cctgaagacc gacgtgaccg actaccacat cgaccaggtg 1980 agcaacctgg tggactgctt aagctagaga tctgttctgc acaaagtgga gtagtcagtc 2040 atcgatcagg aaccagacac cagactttta ttcatacagt gaagtgaagt gaagtgcagt 2100 gcagtgagtt gctggttttt gtaccactta gtatgtattt gtatttgtaa aatacttcta 2160 tcaataaaat ttctaattcc taaaaccaaa atccagtggg taccagcttg ggctgagtgg 2220 ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg 2280 gcgggggtca taacgtgact cccttaattc tccgctcatg atcagattgt cgtttcccgc 2340 cttcagttta aactatcagt gtttgacagg atatattggc gggtaaacct aagagaaaag 2400 agcgtttatt agaataacgg atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat 2460 ttgtatgtgc atgccaacca cagggttccc ctcgggagtg cttggcattc cgtacgataa 2520 tgacttctgt tcaaccaccc aaacgtcgga aagcctgacg acggagcagc attccaaaaa 2580 gatcccttgg ctcgtctggg tcggctagaa ggtcgagtgg gctgctgtgg cttgatccct 2640 caacgcggtc gcggacgtag cgcagcgccg aaaaatcctc gatcgcaaat ccgacgctgt 2700 cgaaaagcgt gatctgcttg tcgctctttc ggccgacgtc ctggccagtc atcacgcgcc 2760 aaagttccgt cacaggatga tctggcgcga gttgctggat ctcgccttca atccgggtct 2820 gtggcgggaa ctccacgaaa atatccgaac gcagcaagat cgtcgaccaa ttcttgaaga 2880 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct 2940 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc 3000 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa 3060 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt 3120 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct 3180 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc 3240 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta 3300 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac 3360 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc 3420 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac 3480 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg 3540 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac 3600 gagcgtgaca ccacgatgcc tgcagggggg gggggggggg ggacatgagg ttgccccgta 3660 ttcagtgtcg ctgatttgta ttgtctgaag ttgtttttac gttaagttga tgcagatcaa 3720 ttaatacgat acctgcgtca taattgatta tttgacgtgg tttgatggcc tccacgcacg 3780 ttgtgatatg tagatgataa tcattatcac tttacgggtc ctttccggtg atccgacagg 3840 ttacggggcg gcgacctcgc gggttttcgc tatttatgaa aattttccgg tttaaggcgt 3900 ttccgttctt cttcgtcata acttaatgtt tttatttaaa ataccctctg aaaagaaagg 3960 aaacgacagg tgctgaaagc gaggcttttt ggcctctgtc gtttcctttc tctgtttttg 4020 tccgtggaat gaacaatgga agtccccccc cccccccccc cctgcagcaa tggcaacaac 4080 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 4140 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 4200 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact 4260 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 4320 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 4380 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 4440 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 4500 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 4560 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 4620 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 4680 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 4740 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 4800 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 4860 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 4920 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 4980 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 5040 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 5100 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 5160 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 5220 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 5280 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt 5340 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg 5400 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg 5460 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 5520 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca 5580 ccgtcatcac cgaaacgcgc gaggcagcag atcccccgat caagtagata cactacatat 5640 atctacaata gacatcgagc cggaaggtga tgtttacttt cctgaaatcc ccagcaattt 5700 taggccagtt tttacccaag acttcgcctc taacataaat tatagttacc aaatctggca 5760 aaagggttaa caagtggcag caacggattc gcaaacctgt cacgcctttt gtgccaaaag 5820 ccgcgccagg tttgcgatcc gctgtgccag gcgttaggcg tcatatgaag atttcggtga 5880 tccctgagca ggtggcggaa acattggatg ctgagaacca tttcattgtt cgtgaagtgt 5940 tcgatgtgca cctatccgac caaggctttg aactatctac cagaagtgtg agcccctacc 6000 ggaaggatta catctcggat gatgactctg atgaagactc tgcttgctat ggcgcattca 6060 tcgaccaaga gcttgtcggg aagattgaac tcaactcaac atggaacgat ctagcctcta 6120 tcgaacacat tgttgtgtcg cacacgcacc gaggcaaagg agtcgcgcac agtctcatcg 6180 aatttgcgaa aaagtgggca ctaagcagac agctccttgg catacgatta gagacacaaa 6240 cgaacaatgt acctgcctgc aatttgtacg caaaatgtgg ctttactctc ggcggcattg 6300 acctgttcac gtataaaact agacctcaag tctcgaacga aacagcgatg tactggtact 6360 ggttctcggg agcacaggat gacgcctaac aattcattca agccgacacc gcttcgcggc 6420 gcggcttaat tcaggagtta aacatcatga gggaagcggt gatcgccgaa gtatcgactc 6480 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac 6540 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg 6600 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg 6660 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg 6720 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat 6780 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg 6840 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg 6900 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc 6960 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc 7020 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg 7080 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctagg caggcttatc 7140 ttggacaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gttcactacg 7200 tgaaaggcga gatcaccaag gtagtcggca aataatgtct aacaattcgt tcaagccgac 7260 gccgcttcgc ggcgcggctt aactcaagcg ttagagagct ggggaagact atgcgcgatc 7320 tgttgaaggt ggttctaagc ctcgtacttg cgatggcatc ggggcaggca cttgctgacc 7380 tgccaattgt tttagtggat gaagctcgtc ttccctatga ctactcccca tccaactacg 7440 acatttctcc aagcaactac gacaactcca taagcaatta cgacaatagt ccatcaaatt 7500 acgacaactc tgagagcaac tacgataata gttcatccaa ttacgacaat agtcgcaacg 7560 gaaatcgtag gcttatatat agcgcaaatg ggtctcgcac tttcgccggc tactacgtca 7620 ttgccaacaa tgggacaacg aacttctttt ccacatctgg caaaaggatg ttctacaccc 7680 caaaaggggg gcgcggcgtc tatggcggca aagatgggag cttctgcggg gcattggtcg 7740 tcataaatgg ccaattttcg cttgccctga cagataacgg cctgaagatc atgtatctaa 7800 gcaactagcc tgctctctaa taaaatgtta ggcctcaaca tctagtcgca agctgagggg 7860 aaccactagt gtcatacgaa cctccaagag acggttacac aaacgggtac attgttgatg 7920 tcatgtatga caatcgccca agtaagtatc cagctgtgtt cagaacgtac gtccgaatta 7980 attcatcggg gtacggtcga cgatcgtcaa cgttcacttc taaagaaata gcgccactca 8040 gcttcctcag cggctttatc cagcgatttc ctattatgtc ggcatagttc tcaagatcga 8100 cagcctgtca cggttaagcg agaaatgaat aagaaggctg ataattcgga tctctgcgag 8160 ggagatgata tttgatcaca ggcagcaacg ctctgtcatc gttacaatca acatgctacc 8220 ctccgcgaga tcatccgtgt ttcaaacccg gcagcttagt tgccgttctt ccgaatagca 8280 tcggtaacat gagcaaagtc tgccgcctta caacggctct cccgctgacg ccgtcccgga 8340 ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt 8400 ggctggctgg tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata 8460 acacattgcg gacgttttta atgtactgaa ttgtctagac ccggggatct catgtttgac 8520 agcttatcat cggatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg 8580 cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa 8640 aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat 8700 tcaacagaaa ttagatgata atcatcgcaa gaccggcaac aggattcaat cttaagaaac 8760 tttattgcca aatgtttgaa cgatctctgc aggtcgacgg atcgagctcc cagcttagca 8820 agagatgtta attttttcag taagctctta cagcttgttg taaacacgcg ctaaacggcc 8880 gtggcctttg acagtcaccg gtgattcgtt ggcggcaata aacgctgatt cacccggttt 8940 aagctgtaac tgctgagaac ctttccacaa cgttgcatcg ccttcgacgc agaacaaaat 9000 ggcggcactc tgctggctaa tggtggtttc tttatcacta aggtcatgca gcgagaaggc 9060 aaaatcatcc actggaatcg ggaagtccag ttctgcacct tgtttcaccg gctgggtcaa 9120 caactggtta gccggtttgg cttcgaattt cacattggca accagttccg gaatatcaat 9180 gtatttaggc gtcagacccg cacgcagcac gttatcggag tttgccatca cttccagcgc 9240 cacgccttgc aggtaagcgt gcggtgtttc agcgaacagg aacatcgctt cgccagggtt 9300 caatttcacc acattcagca atagcgggga gaacagaccg ctgtcttccg ggtaaaattc 9360 agaaattaaa cgaatcgttt gccacggttc accctgctgg ctatcgaggg ccgattttaa 9420 aatcgccagc gcgcgggatt tttcttcacc ctgcatattc aacaggctgg cgaacagttc 9480 gcttaaacgt tcggcatcag gctgttgtaa aaagtgagca atcgccggat gtgcacctgc 9540 gaccggctgg agtagggaga caatctcgga aaattcacga aacgcgttca tcgcaaggaa 9600 aggcgtcagc gcaaaaacca gctccggctt gtggttagga tctttatagt tacgctcggc 9660 ggcatccatc gggatacctg cggcattttc tttggcaaaa ccgatttcag aattgtgttt 9720 gtttggatga acctgaatgg agagtggctg tgctgcgcat aatactttga acaggaaagg 9780 cagttcgcca aagcgtttgg caacggcctc tccgagcaga gtcgatttat cactctcaat 9840 cacatcacgc agtgaaacga tatctccggc ggcattctgc actcgtgaac tgcttttcgg 9900 atgtgcgccc atccacagct cggccatcgg ctggctggac ggattttcca taccataaag 9960 ttcagtcaac gcgttttgct gccccaggca tagttttgca ctgagttaat gagtttttgc 10020 atgatcgggg atccctgcag aagtaacacc aaacaacagg gtgagcatcg acaaaagaaa 10080 cagtaccaag caaataaata gcgtatgaag gcagggctaa aaaaatccac atatagctgc 10140 tgcatatgcc atcatccaag tatatcaaga tcaaaataat tataaaacat acttgtttat 10200 tataatagat aggtactcaa ggttagagca tatgaataga tgctgcatat gccatcatgt 10260 atatgcatca gtaaaaccca catcaacatg tatacctatc ctagatcgat atttccatcc 10320 atcttaaact cgtaactatg aagatgtatg acacacacat acagttccaa aattaataaa 10380 tacaccaggt agtttgaaac ggcgtctact ccgatctaga acgaatgaac gaccgcccaa 10440 ccacaccaca tcatcacaac caagcgaaca aaaagcatct ctgtatatgc atcagtaaaa 10500 cccgcatcaa catgtatacc tatcctagat cgatatttcc atccatcatc ttcaattcgt 10560 aactatgaat atgtatggca cacacataca gatccaaaat taataaatcc accaggtagt 10620 ttgaaacaga attctactcc gatctagaac gaccgcccaa ccagaccaca tcatcacaac 10680 caagacaaaa aaaagcatga aaagatgacc cgacaaacaa gtgcacggca tatattgaaa 10740 taaaggaaaa gggcaaacca aaccctatgc aacgaaacaa aaaaaatcat gaaatcgatc 10800 ccgtctgcgg aacggctaga gccatcccag gattccccaa agagaaacac tggcaagtta 10860 gcaatcagaa cgtgtctgac gtacaggtcg catccgtgta cgaacgctag cagcacggat 10920 ctaacacaaa cacggatcta acacaaacat gaacagaagt agaactaccg ggccctaacc 10980 atggaccgga acgccgatct agagaaggta gagagggggg gggggggagg acgagcggcg 11040 taccttgaag cggaggtgcc gacgggtgga tttgggggag atctggttgt gtgtgtgtgc 11100 gctccgaaca acacgaggtt ggggaaagag ggtgtggagg gggtgtctat ttattacggc 11160 gggcgaggaa gggaaagcga aggagcggtg ggaaaggaat cccccgtagc tgccgtgccg 11220 tgagaggagg aggaggccgc ctgccgtgcc ggctcacgtc tgccgctccg ccacgcaatt 11280 tctggatgcc gacagcggag caagtccaac ggtggagcgg aactctcgag aggggtccag 11340 aggcagcgac agagatgccg tgccgtctgc ttcgcttggc ccgacgcgac gctgctggtt 11400 cgctggttgg tgtccgttag actcgtcgac ggcgtttaac aggctggcat tatctactcg 11460 aaacaagaaa aatgtttcct tagttttttt aatttcttaa agggtatttg tttaattttt 11520 agtcacttta ttttattcta ttttatatct aaattattaa ataaaaaaac taaaatagag 11580 ttttagtttt cttaatttag aggctaaaat agaataaaat agatgtacta aaaaaattag 11640 tctataaaaa ccattaaccc taaaccctaa atggatgtac taataaaatg gatgaagtat 11700 tatataggtg aagctatttg caaaaaaaaa ggagaacaca tgcacactaa aaagataaaa 11760 ctgtagagtc ctgttgtcaa aatactcaat tgtcctttag accatgtcta actgttcatt 11820 tatatgattc tctaaaacac tgatattatt gtagtactat agattatatt attcgtagag 11880 taaagtttaa atatatgtat aaagatagat aaactgcact tcaaacaagt gtgacaaaaa 11940 aaatatgtgg taatttttta taacttagac atgcaatgct cattatctct agagaggggc 12000 acgaccgggt cacgctgcac tgcaggcatg caagcttgca catgacaaca attgtaagag 12060 gatggagacc acaacgatcc aacaatactt ctgcgacggg ctgtgaagta tagagaagtt 12120 aaacgcccaa aagccattgt gtttggaatt tttagttatt ctatttttca tgatgtatct 12180 tcctctaaca tgccttaatt tgcaaatttg gtataactac tgattgaaaa tatatgtatg 12240 taaaaaaata ctaagcatat ttgtgaagct aaacatgatg ttatttaaga aaatatgttg 12300 ttaacagaat aagattaata tcgaaatgga aacatctgta aattagaatc atcttacaag 12360 ctaagagatg ttcacgcttt gagaaacttc ttcagatcat gaccgtagaa gtagctctcc 12420 aagactcaac gaaggctgct gcaattccac aaatgcatga catgcatcct tgtaaccgtc 12480 gtcgccgcta taaacacgga taactcaatt ccctgctcca tcaatttaga aatgagcaag 12540 caagcacccg atcgctcacc ccatatgcac caatctgact cccaagtctc tgtttcgcat 12600 tagtaccgcc agcactccac ctatagctac caattgagac ctttccagcc taagcagatc 12660 gattgatcgt tagagtcaaa gagttggtgg tacgggtact ttaactacca tggaatgatg 12720 gggcgtgatg tagagcggaa agcgcctccc tacgcggaac aacaccctcg ccatgccgct 12780 cgactacagc ctcctcctcg tcggccgccc acaacgaggg agcccgtggt cgcagccacc 12840 gaccagcatg tctctgtgtc ctcgtccgac ctcgacatgt catggcaaac agtcggacgc 12900 cagcaccaga ctgacgacat gagtctctga agagcccgcc acctagaaag atccgagccc 12960 tgctgctggt agtggtaacc attttcgtcg cgctgacgcg gagagcgaga ggccagaaat 13020 ttatagcgac tgacgctgtg gcaggcacgc tatcggaggt tacgacgtgg cgggtcactc 13080 gacgcggagt tcacaggtcc tatccttgca tcgctcgggc cggagtttac gggacttatc 13140 cttacgacgt gctctaaggt tgcgataacg ggcggaggaa ggcgtgtggc gtgcggagac 13200 ggtttataca cgtagtgtgc gggagtgtgt ttcgtagacg cgggaaagca cgacgactta 13260 cgaaggttag tggaggagga ggacacacta aaatcaggac gcaagaaact cttctattat 13320 agtagtagag aagagattat aggagtgtgg gttgattcta aagaaaatcg acgcaggaca 13380 accgtcaaaa cgggtgcttt aatatagtag atatatatat atagagagag agagaaagta 13440 caaaggatgc atttgtgtct gcatatgatc ggagtattac taacggccgt cgtaagaagg 13500 tccatcatgc gtggagcgag cccatttggt tggttgtcag gccgcagtta aggcctccat 13560 atatgattgt cgtcgggccc ataacagcat ctcctccacc agtttattgt aagaataaat 13620 taagtagaga tatttgtcgt cgggcagaag aaacttggac aagaagaaga agcaagctag 13680 gccaatttct tgccggcaag aggaagatag tggcctctag tttatatatc ggcgtgatga 13740 tgatgctcct agctagaaat gagagaagaa aaacggacgc gtgtttggtg tgtgtcaatg 13800 gcgtccatcc ttccatcaga tcagaacgat gaaaaagtca agcacggcat gcatagtata 13860 tgtatagctt gttttagtgt ggctttgctg agacgaatga aagcaacggc gggcatattt 13920 ttcagtggct gtagctttca ggctgaaaga gacgtggcat gcaataattc agggaattcg 13980 tcagccaatt gaggtagcta gtcaacttgt acattggtgc gagcaatttt ccgcactcag 14040 gagggctagt ttgagagtcc aaaaactata ggagattaaa gaggctaaaa tcctctcctt 14100 atttaatttt aaataagtag tgtatttgta ttttaactcc tccaaccctt ccgattttat 14160 ggctctcaaa ctagcattca gtctaatgca tgcatgcttg gctagaggtc gtatggggtt 14220 gttaatagca tagctagcta caagttaacc gggtctttta tatttaataa ggacaggcaa 14280 agtattactt acaaataaag aataaagcta ggacgaactc gtggattatt actaaatcga 14340 aatggacgta atattccagg caagaataat tgttcgatca ggagacaagt ggggcattgg 14400 accggttctt gcaagcaaga gcctatggcg tggtgacacg gcgcgttgcc catacatcat 14460 gcctccatcg atgatccatc ctcacttgct ataaaaagag gtgtccatgg tgctcaagct 14520 cagccaagca aataagacga cttgtttcat tgattcttca agagatcgag cttcttttgc 14580 accacaaggt cgaggatcca aca 14603 <210> 17 <211> 11127 <212> DNA <213> Artificial Sequence <220> <223> Description of Artificial Sequence: pZU578 <220> <221> misc_feature (222) (1485) .. (3491) <223> synthetic nucleotide sequence encoding the toxin       portion of H04 plus the first 40 amino acids of       the Cry1Ab tail <220> <221> misc_feature (222) (5052) .. (6271) <223> PMI <220> <221> misc_feature (222) (3859) .. (5030) <223> SMAS promoter <220> <221> misc_feature (222) (56) .. (1475) <223> Actin 2 promoter U41998 <400> 17 ggccgcagcg gccatttaaa tcaattgggc gcgccgaatt cgagctcggt accctgcatg 60 cctgcaggtc gacaaaattt agaacgaact taattatgat ctcaaataca ttgatacata 120 tctcatctag atctaggtta tcattatgta agaaagtttt gacgaatatg gcacgacaaa 180 atggctagac tcgatgtaat tggtatctca actcaacatt atacttatac caaacattag 240 ttagacaaaa tttaaacaac tattttttat gtatgcaaga gtcagcatat gtataattga 300 ttcagaatcg ttttgacgag ttcggatgta gtagtagcca ttatttaatg tacatactaa 360 tcgtgaatag tgaatatgat gaagcattgt atcttattgt ataaatatcc ataaacacat 420 catgaaagac actttctttc acggtctgaa ttaattatga cacaattcta atagaaaacg 480 aattaaatta cgttgaattg tatgaaatct aattgaacaa gccaaccacg acgacgacta 540 acgttgcctg gattgactcg gtttaagtta accactaaaa aaacggagct gtcatgtaac 600 acgcggatcg agcaggtcac agtcatgaag ccatcaaagc aaaagaacta atccaagggc 660 tgagatgatt aattagttta aaaattagtt aacacgaggg aaaaggctgt ctgacagcca 720 ggtcacgtta tctttacctg tggtcgaaat gattcgtgtc tgtcgatttt aattattttt 780 ttgaaaggcc gaaaataaag ttgtaagaga taaacccgcc tatataaatt catatatttt 840 cctctccgct ttgaattgtc tcgttgtcct cctcactttc atcagccgtt ttgaatctcc 900 ggcgacttga cagagaagaa caaggaagaa gactaagaga gaaagtaaga gataatccag 960 gagattcatt ctccgttttg aatcttcctc aatctcatct tcttccgctc tttctttcca 1020 aggtaatagg aactttctgg atctacttta tttgctggat ctcgatcttg ttttctcaat 1080 ttccttgaga tctggaattc gtttaatttg gatctgtgaa cctccactaa atcttttggt 1140 tttactagaa tcgatctaag ttgaccgatc agttagctcg attatagcta ccagaatttg 1200 gcttgacctt gatggagaga tccatgttca tgttacctgg gaaatgattt gtatatgtga 1260 attgaaatct gaactgttga agttagattg aatctgaaca ctgtcaatgt tagattgaat 1320 ctgaacactg tttaagttag atgaagtttg tgtatagatt cttcgaaact ttaggatttg 1380 tagtgtcgta cgttgaacag aaagctattt ctgattcaat cagggtttat ttgactgtat 1440 tgaactcttt ttgtgtgttt gcagctcata aaaaggatcc aacaatggac aacaacccca 1500 acatcaacga gtgcatcccc tacaactgcc tgagcaaccc cgaggtggag gtgctgggcg 1560 gcgagcgcat cgagaccggc tacaccccca tcgacatcag cctgagcctg acccagttcc 1620 tgctgagcga gttcgtgccc ggcgccggct tcgtgctggg cctggtggac atcatctggg 1680 gcatcttcgg ccccagccag tgggacgcct tcctggtgca gatcgagcag ttgataaacc 1740 aacgcataga ggaattcgcc cgcaaccagg ccatcagccg cctggagggc ctgagcaacc 1800 tgtaccaaat ctacgccgag agcttccgcg agtgggaggc cgaccccacc aaccccgccc 1860 tgcgcgagga gatgcgcatc cagttcaacg acatgaacag cgccctgacc accgccatcc 1920 ccctgttcgc cgtgcagaac taccaggtgc ccctgctgag cgtgtacgtg caggccgcca 1980 acctgcacct gagcgtgctg cgcgacgtca gcgtgttcgg ccagcgctgg ggcttcgacg 2040 ccgccaccat caacagccgc tacaacgacc tgacccgcct gatcggcaac tacaccgacc 2100 acgccgtgcg ctggtacaac accggcctgg agcgcgtgtg gggtcccgac agccgcgact 2160 ggatcaggta caaccagttc cgccgcgagc tgaccctgac cgtgctggac atcgtgagcc 2220 tgttccccaa ctacgacagc cgcacctacc ccatccgcac cgtgagccag ctgacccgcg 2280 agatttacac caaccccgtg ctggagaact tcgacggcag cttccgcggc agcgcccagg 2340 gcatcgaggg cagcatccgc agcccccacc tgatggacat cctgaacagc atcaccatct 2400 acaccgacgc ccaccgcggc gagtactact ggagcggcca ccagatcatg gccagccccg 2460 tcggcttcag cggccccgag ttcaccttcc ccctgtacgg caccatgggc aacgctgcac 2520 ctcagcagcg catcgtggca cagctgggcc agggagtgta ccgcaccctg agcagcaccc 2580 tgtaccgtcg acctttcaac atcggcatca acaaccagca gctgagcgtg ctggacggca 2640 ccgagttcgc ctacggcacc agcagcaacc tgcccagcgc cgtgtaccgc aagagcggca 2700 ccgtggacag cctggacgag atcccccctc agaacaacaa cgtgccacct cgacagggct 2760 tcagccaccg tctgagccac gtgagcatgt tccgcagtgg cttcagcaac agcagcgtga 2820 gcatcatccg tgcacccatg ttcagctgga ttcaccgcag cgccaccctg accaacacca 2880 tcgaccccga gcgcatcaac cagatccccc tggtgaaggg cttccgggtg tggggcggca 2940 ccagcgtgat caccggcccc ggcttcaccg gaggcgacat cctgcgcaga aacaccttcg 3000 gcgacttcgt gagcctgcag gtgaacatca acagccccat cacccagcgt taccgcctgc 3060 gcttccgcta cgccagcagc cgcgacgccc gtgtgatcgt gctgactggc gccgctagca 3120 ccggtgtggg cggtcaggtg agcgtgaaca tgcccctgca gaagactatg gagatcggcg 3180 agaacctgac tagtcgcacc ttccgctaca ccgacttcag caaccccttc agcttccgcg 3240 ccaaccccga catcatcggc atcagcgagc agcccctgtt cggtgccggc agcatcagca 3300 gcggcgagct gtacatcgac aagatcgaga tcatcctggc cgacgccacc ttcgaggccg 3360 agagcgacct ggagcgcgcc cagaaggccg tgaacgccct gttcaccagc agcaaccaga 3420 tcggcctgaa gaccgacgtg accgactacc acatcgacca ggtgagcaac ctggtggact 3480 gcttaagcta gagatcctct agagtcgacc atggtgatca ctgcagatcg ttcaaacatt 3540 tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa 3600 tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg 3660 agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa 3720 atatagcgcg caacctagga taaattatcg cgcgcggtgt catctatgtt actagatctc 3780 tagaaagctt cgtacgttaa ttaattcgaa tccggagcgg ccgcagggct agcatcgatg 3840 gtaccgagct cgagactata caggccaaat tcgctcttag ccgtacaata ttactcaccg 3900 gtgcgatgcc ccccatcgta ggtgaaggtg gaaattaatg atccatcttg agaccacagg 3960 cccacaacag ctaccagttt cctcaagggt ccaccaaaaa cgtaagcgct tacgtacatg 4020 gtcgataaga aaaggcaatt tgtagatgtt aacatccaac gtcgctttca gggatcccga 4080 attccaagct tggaattcgg gatcctacag gccaaattcg ctcttagccg tacaatatta 4140 ctcaccggtg cgatgccccc catcgtaggt gaaggtggaa attaatgatc catcttgaga 4200 ccacaggccc acaacagcta ccagtttcct caagggtcca ccaaaaacgt aagcgcttac 4260 gtacatggtc gataagaaaa ggcaatttgt agatgttaac atccaacgtc gctttcaggg 4320 atcccgaatt ccaagcttgg aattcgggat cctacaggcc aaattcgctc ttagccgtac 4380 aatattactc accggtgcga tccccccatc gtaggtgaag gtggaaatta atgatccatc 4440 ttgagaccac aggcccacaa cagctaccag tttcctcaag ggtccaccaa aaacgtaagc 4500 gcttacgtac atggtcgata agaaaaggca atttgtagat gttaacatcc aacgtcgctt 4560 tcagggatcc cgaattccaa gcttgggctg caggtcaatc ccattgcttt tgaagcagct 4620 caacattgat ctctttctcg agggagattt ttcaaatcag tgcgcaagac gtgacgtaag 4680 tatccgagtc agtttttatt tttctactaa tttggtcgtt tatttcggcg tgtaggacat 4740 ggcaaccggg cctgaatttc gcgggtattc tgtttctatt ccaacttttt cttgatccgc 4800 agccattaac gacttttgaa tagatacgct gacacgccaa gcctcgctag tcaaaagtgt 4860 accaaacaac gctttacagc aagaacggaa tgcgcgtgac gctcgcggtg acgccatttc 4920 gccttttcag aaatggataa atagccttgc ttcctattat atcttcccaa attaccaata 4980 cattacacta gcatctgaat ttcataacca atctcgatac accaaatcga gatctgcagg 5040 gatccccgat catgcaaaaa ctcattaact cagtgcaaaa ctatgcctgg ggcagcaaaa 5100 cggcgttgac tgaactttat ggtatggaaa atccgtccag ccagccgatg gccgagctgt 5160 ggatgggcgc acatccgaaa agcagttcac gagtgcagaa tgccgccgga gatatcgttt 5220 cactgcgtga tgtgattgag agtgataaat cgactctgct cggagaggcc gttgccaaac 5280 gctttggcga actgcctttc ctgttcaaag tattatgcgc agcacagcca ctctccattc 5340 aggttcatcc aaacaaacac aattctgaaa tcggttttgc caaagaaaat gccgcaggta 5400 tcccgatgga tgccgccgag cgtaactata aagatcctaa ccacaagccg gagctggttt 5460 ttgcgctgac gcctttcctt gcgatgaacg cgtttcgtga attttccgag attgtctccc 5520 tactccagcc ggtcgcaggt gcacatccgg cgattgctca ctttttacaa cagcctgatg 5580 ccgaacgttt aagcgaactg ttcgccagcc tgttgaatat gcagggtgaa gaaaaatccc 5640 gcgcgctggc gattttaaaa tcggccctcg atagccagca gggtgaaccg tggcaaacga 5700 ttcgtttaat ttctgaattt tacccggaag acagcggtct gttctccccg ctattgctga 5760 atgtggtgaa attgaaccct ggcgaagcga tgttcctgtt cgctgaaaca ccgcacgctt 5820 acctgcaagg cgtggcgctg gaagtgatgg caaactccga taacgtgctg cgtgcgggtc 5880 tgacgcctaa atacattgat attccggaac tggttgccaa tgtgaaattc gaagccaaac 5940 cggctaacca gttgttgacc cagccggtga aacaaggtgc agaactggac ttcccgattc 6000 cagtggatga ttttgccttc tcgctgcatg accttagtga taaagaaacc accattagcc 6060 agcagagtgc cgccattttg ttctgcgtcg aaggcgatgc aacgttgtgg aaaggttctc 6120 agcagttaca gcttaaaccg ggtgaatcag cgtttattgc cgccaacgaa tcaccggtga 6180 ctgtcaaagg ccacggccgt ttagcgcgtg tttacaacaa gctgtaagag cttactgaaa 6240 aaattaacat ctcttgctaa gctgggagct cgtcgacgga tcgaattcct gcagatcgtt 6300 caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta 6360 tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt 6420 tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag 6480 aaaacaaaat atagcgcgca acctaggata aattatcgcg cgcggtgtca tctatgttac 6540 tagatctcta gaactagtgg atctgctagc cctgcaggaa atttaccggt gcccgggcgg 6600 ccagcatggc cgtatccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc 6660 acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc 6720 accactcgat acaggcagcc catcagaatt aattctcatg tttgacagct tatcatcgac 6780 tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg 6840 caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg 6900 ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt 6960 aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca 7020 gaccatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt 7080 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga 7140 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga 7200 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga 7260 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg 7320 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc 7380 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag 7440 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga 7500 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg 7560 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac 7620 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca 7680 gtatcagccc gtcatacttg aagctaggca ggcttatctt ggacaagaag atcgcttggc 7740 ctcgcgcgca gatcagttgg aagaatttgt tcactacgtg aaaggcgaga tcaccaaggt 7800 agtcggcaaa taaagctcta gtggatcccc gaggaatcgg cgtgacggtc gcaaaccatc 7860 cggcccggta caaatcggcg cggcgctggg tgatgacctg gtggagaagt tgaaggccgc 7920 gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc 7980 ggccgctgat cgaatccgca aagaatcccg gcaaccgccg gcagccggtg cgccgtcgat 8040 taggaagccg cccaagggcg acgagcaacc agattttttc gttccgatgc tctatgacgt 8100 gggcacccgc gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga 8160 ccgacgagct ggcgaggtga tccgctacga gcttccagac gggcacgtag aggtttcagc 8220 agggccggcc ggcatggcca gtgtgtggga ttacgacctg gtactgatgg cggtttccca 8280 tctaaccgaa tccatgaacc gataccggga agggaaggga gacaagcccg gccgcgtgtt 8340 ccgtccacac gttgcggacg tactcaagtt ctgccggcga gccgatggcg gaaagcagaa 8400 agacgacctg gtagaaacct gcattcggtt aaacaccacg cacgttgcca tgcagcgtac 8460 gaagaaggcc aagaacggcc gcctggtgac ggtatccgag ggtgaagcct tgattagccg 8520 ctacaagatc gtaaagagcg aaaccgggcg gccggagtac atcgagatcg agctagctga 8580 ttggatgtac cgcgagatca cagaaggcaa gaacccggac gtgctgacgg ttcaccccga 8640 ttactttttg atcgatcccg gcatcggccg ttttctctac cgcctggcac gccgcgccgc 8700 aggcaaggca gaagccagat ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg 8760 agagttcaag aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg acctgccgga 8820 gtacgatttg aaggaggagg cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa 8880 cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc tagggcaaat 8940 tgccctagca ggggaaaaag gtcgaaaagg tctctttcct gtggatagca cgtacattgg 9000 gaacccaaag ccgtacattg ggaaccggaa cccgtacatt gggaacccaa agccgtacat 9060 tgggaaccgg tcacacatgt aagtgactga tataaaagag aaaaaaggcg atttttccgc 9120 ctaaaactct ttaaaactta ttaaaactct taaaacccgc ctggcctgtg cataactgtc 9180 tggccagcgc acagccgaag agctgcaaaa agcgcctacc cttcggtcgc tgcgctccct 9240 acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa tggctggcct 9300 acggccaggc aatctaccag ggcgcggaca agccgcgccg tcgccactcg accgccggcg 9360 ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggcctgaa tcgccccatc 9420 atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg tggaccagtt 9480 ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat 9540 ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccgccgtc ccgtcaagtc 9600 agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg 9660 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa 9720 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc 9780 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg 9840 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat 9900 ggcaaaagct ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 9960 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 10020 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 10080 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 10140 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 10200 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 10260 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 10320 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 10380 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 10440 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 10500 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 10560 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 10620 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 10680 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 10740 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 10800 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttgatcc ggaattaatt 10860 cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac gcccttttaa 10920 atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa tatatcctgt 10980 caaacactga tagtttaaac tgaaggcggg aaacgacaat ctgatcatga gcggagaatt 11040 aagggagtca cgttatgacc cccgccgatg acgcgggaca agccgtttta cgtttggaac 11100 tgacagaacc gcaacgctgc aggaatt 11127

Claims (59)

delete delete delete delete delete delete delete delete delete A synthetic nucleotide sequence encoding a hybrid Bacillus thuringiensis toxin, selected from the group consisting of SEQ ID NOs: 5, 7 and 9. delete The synthetic nucleotide sequence of claim 10, wherein said hybrid toxin has an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 8, and 10. 12. delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete An isolated nucleic acid molecule encoding a hybrid Bacillus thuringiensis toxin having a nucleotide sequence selected from the group consisting of SEQ ID NOs: 5, 7, 9, 12, 13, 14, 15, 16, and 17. delete delete A chimeric gene comprising the synthetic nucleotide sequence of claim 10 or 12 or a heterologous promoter sequence operably linked to the nucleic acid molecule of claim 43. A recombinant vector comprising the chimeric gene of claim 46. delete A transgenic plant cell comprising the chimeric gene of claim 46 or the recombinant vector of claim 47. delete 50. The transgenic plant cell of claim 49, which is derived from corn, cotton, rice or cabbage. delete Fall armyworms, pink bollworms, tobacco budworms, European moths A method for controlling pests selected from the group consisting of European corn borer) and diamondback moth. 54. The method of claim 53, wherein the transgenic plant is corn, cotton, rice or cabbage. A method for producing insect-resistant plants capable of controlling pests, comprising introducing a synthetic nucleotide sequence according to claim 10 into a plant. 56. The method of claim 55, wherein the pest is an insect of the species. 57. The method of claim 56, wherein the pest is chestnut moth, pink cotton worm, tobacco moth, European moth or cabbage moth. The method of claim 55, wherein the plant is corn, cotton, rice or cabbage. delete